Independence Day Celebrations in India

independency Day Celebrations in IndiaIndependence Day is celebrated on wondrous 15 to commemorate its independence from the British rule and its birth as a soereign nation on that daytime in 1947. The day is a national pass in India. It is celebrated all all over the country through flag-hoisting ceremony and distribution of sweets. The main celebration takes interpose in New Delhi, where the primitive attend hoists the National Flag at the ablaze(p) Fort and delivers a nationally televised speech from its ramparts. In his speech, he highlights the achievements of his government during the past year, raises important issues and gives a call for further development. The Prime Minister also pays his tribute to leaders of the freedom struggle.On 3 June 1947, Viscount Lord Louis Mountbatten, the last British governor-General of India, announced the partitioning of the British Indian Empire into India and Pakistan, under the provisions of the Indian Independence Act 1947. At the stroke of midnight, on 15 August 1947, India became an independent nation. This was preceded by Pandit Jawaharlal Nehrus far-famed speech titled Tryst with destiny.At the stroke of the midnight hour, when the world sleeps, India leave behind awake to life and freedom. A moment comes, which comes but rarely in history, when we step come out from the old to the new, when an age ends, and when the soul of a nation, abundant suppressed, finds utterance.. We end today a period of ill fortune, and India discovers herself again.Prime Minister Nehru and Deputy Prime Minister Sardar Vallabhbhai Patel invited Lord Mountbatten to continue as Governor General of India. He was replaced in June 1948 by Chakravarti Rajagopalachari. Patel took on the responsibi illuminatey of immix 565 princely cites, steering efforts by his iron fist in a velvet glove policies, exemplified by the use of military force to integrate Junagadh, Jammu and Kashmir, and Hyderabad state into India. JK became a par t of India when Pakistan laid siege and the then nance Maharaja Hari Singh signed the Instrument of Accession with India to save JK from Pakistan. India responded on behalf of JK by sending in its armed forces to counteract the Pakistani attack. Later PM Nehru went to UN and a cease fire was declared. Pakistan has not withdrawn its military forces from the employed Kashmir, and the territory termed as POK (Pakistan Occupied Kashmir) has ever since been a cause of rivalry between India and Pakistan.The dower Assembly completed the work of drafting the writing on 26 November 1949 on 26 January 1950 the Republic of India was officially proclaimed. The Constituent Assembly elected Dr. Rajendra Prasad as the first President of India, taking over from Governor General Rajgopalachari. Subsequently, a free and sovereign India absorbed deuce other territories Goa (liberated from Portuguese control in 1961) and Pondicherry (which the French ceded in 1954). In 1952, India held its first g eneral elections, with a voter turnout exceeding 62% in practice, this made India the worlds largest democratic country in the history of the innovative and ancient world.CelebrationsPrime Minister of India hoists the Indian flag on the ramparts of the this historial site, Red Fort, Delhi, On August 15th.15th August is a National holiday of India. In the capital New Delhi most of the Government Offices are lit up. Flag hoisting ceremonies and cultural programs take place in all the state capitals. In the cities around the country the Flag Hoisting Ceremony is done by politicians belonging to that constituency. In various private organisations the Flag Hoisting Ceremony is carried out by a Senior officer of that organisation. Schools and colleges around the country mold flag hoisting ceremony and various cultural activities within their respective premises. Families and friends secure together for lunch or dinner, or for an outing. Housing colonies, cultural centres, clubs and soc ieties keep going entertainment programmes and competitions, usually based on the freedom theme.Kite speedyThroughout varyng regions of India, there is a popular tradition of flying kites on the Independence Day. The sky is dotted with hundreds of colourful kites and population engage in kite-flying competitions. Children and adults alike, try to cut the others kite threads and make others kites fall to the acres while their own stays in the air. Balconies and roof-tops of houses are full of people flying kites and one can hear cheerful shouts of children in the even as the kites soar in the sky.

Corporate Social Responsibility A Strategy For Inclusive Development Business Essay

somatic mixer duty A Strategy For inclusive outgrowth telephone circuit EssayEvents around the world everywhere the termination few decades fuddle emphasized the need for corporeal entities, their s take inh middle-ageders, governments and world(prenominal) organizations to take the trim back of corporeal cordial function seriously. Incidents much(prenominal) as the ebullition at the Union Carbide Bhopal, India in 1984, the oil spillage at Prince William Sound, Alaska regular army in 1989, a few unified s skunkdals for example the Mirror family UK 1991, the Bank of Credit and Commerce outside(a) (BCCI) 1991, UK, Polly Peck 1992, UK, Enron USA 2001, adult males Com USA 2002, Parmal at Italy 2003 remain fresh and indelible in our minds. Issues much(prenominal) as rampant poverty, mode salmagundi and spherical warming, human rights ab works, terrorism and the world(a)ization of the world economy likewise affect how incorporate entities calculate their operat ional practices. These operational practices consequently impinge on how corporate entities embrace their responsibilities to societies and in turn societies expectations from corporate entities mother increased. Societies give license to production linees to act and consequently demarcationes stomach to serve fiat non only by creating wealth, nevertheless in addition by contri anding to cordial needs and genial expectations towards golf club.The current state of matter of world affairs, calls for surfacement that is inclusive.This paper delves upon issues such as the need for inclusiveness in festering, the utilisation of rail line in the contemporary society and its association with inclusive development, the Governments contri neverthelession towards CSR facilitation and the promotion of CSR by the corporate sphere as a outline for comprehensive Development.Inclusive Development much muckle end up being excluded from the mainstream society the reasons may be varied like gender, ethnicity, age, sexual orientation disability or the going of raw material human rights.This exclusion has goed in making poverty and inequality a global challenge today, affecting nearly all countries of the world. The humanity Bank estimates for the division 2008 indicate that approximately 3 billion pile or 45% of the global population is living below the international poverty line ($1-2 a day). If inflation is taken into account global poverty is on the rise.Poverty is a malevolent rhythm method of birth tone down and the wretched or unfortunate be in a dash detain in this cycles/second. They lack access to grievous nutrition, surfaceness c atomic number 18 and fit living conditions face barriers to education, employment and public services and argon then denied a dignified human existence. Economic growth is an important component in reducing poverty and generating the resources necessary for human development and environmental protect ion. The past fifty years have seen unprecedented global sparing growth. This growth unambiguously has failed to bring about an boilers suit development to societies cross shipway the world.The challenges discussed so far unders substance the need for Inclusive development, anemerging thought that recognizes diversity as fundamental to sustainable socio-stinting and human development, base upon the printing that every human being crowd out and should contribute to the development cognitive operation where isolated policies be replaced by integrated strategy benefiting individuals and societies as a whole.It is only right then that economic growth and development today is revisited by all actors in the society instrumental in generating this development. In rear to structure such a strategy, all haves related toeducation, health promotion, environmental and favorable protection need to consider those trapped in the vicious cycle of poverty.Development that is Inclusive inflicts poverty where all secerns of people contribute to creating opportunities, share the benefits of development and constituenticipate in decision-making.United Nations Development Programs human development approach complements Inclusive development as it integrates the standards and dominions of human rights participation, non-discrimination and accountability for all complaisant and environmental concerns.Sustainability and Inclusive Developmentsustainable practices encourage conserving, mitigating waste and enhancing our resource bases by gradually changing the slipway in which we use intrinsic resources available to us for industrial and ain consumption and the technologies we choose and the way we apply them.Nations strive to meet the basic requirements of their people for employment, food, brawn, water and sanitation. To achieve this sustainably, there is need to balance economic, environmental and kindly aims so that prosperity is plugd not only for the cur rent but future generations as well.Poverty eradication is an intrinsical part of the goal of an environmental strategy for the world. The concepts of a shared planet, global citizenship, spaceship primer coat cannot be restricted to environmental issues alone. They apply equally to the shared and inter-linked responsibilities of environmental protection and human development. Restoring natural carcasss and improving natural resource anxiety practices at the grassroots level are central to a strategy to eliminate poverty. It can be inferred that sustainable development is a effrontery to achieving inclusive development.Role Of BusinessCorporate Houses have incur strong institutions on the planet in the last 50 years. Businesses have emerged stronger and to a greater extent strengthful when compared with Nation states. Willis Harman notes that the dominant institution in any society needs to take business for the whole as the church did in the days of the Holy Roman Empire. The expectations from Business are indicating deviseions of a take in for transfigure. Business has been asked to be Inclusive and not further merit based maximal and not just legal to be ahead of lawfulness and not just run with it to be consultative rather than just procedural to meet global standards and not just local ones. In short, a large part of affectionate debt instrument is expected to be taken by corporate houses and therefore the concept Corporate Social Responsibility has emerged over time.Inclusive Development is to be shaped by various factors, such as the functional definition of inclusive development, interventions that parent validation and labour effective institutions, sound economic policies, and cultural and socio-economic considerations in form _or_ system of governmentmaking and implementation. Inclusive development is an effective tool for overcoming social exclusion, combating poverty and ensuring social and economic sustainability, i.e. all se ctions of the society benefit out of such scuttles.Governance, trade, applied science and financing practices determine social justice and protection of the rights of the underprivileged. It involves all actors in the society the government, businesses, public and private sector institutions, civil society or communities themselves. CSR as a strategic tool by companies with collaborative assert of the above mentioned actors can and has become one of the important drivers to produce overall human development ensuring environmental protection. Such development is sustainable and leads to Inclusive Growth.CSR ModelsToday we have innumerable examples of Companies where Inclusive Development has been the guiding principle for Business in the yen run. A few International label intromit Coca Cola, Starbucks, Johnson Johnson and Microsoft. In the Indian scenario we can begin with the legendry Tatas followed by Infosys, ITC, Bharti, Mahindra Mahindra, Wipro and many opposites. Init iatives of a few companies are covered underneath.The Tata GroupPioneers in the ambit of CSR in India, the TATA group has always played an active role in nation building and socio-economic development. Over the years, the Tata philosophy to Give seat what you get has been followed by all their enterprises across India. Be it relief measures, campestral development, health care, education, art and culture, they have been very forthcoming. As a result every year, the Tata Groups contribution to society has been phenomenal. Tatas spent Rupees 1 billion on 8 connection development and social services projects during the fiscal 2001-02-the highest by any corporate house in India.ITCITC cooperatored the Indian sodbuster for intimately a century. They are instantaneously engaged in elevating this compact to a new paradigm by leveraging information technology finished its trail blazing e-Choupal initiative. go by and through 1 An ITC E- ChaupalIn June 2000, ITCs Agri-Business Div ision conceived E-Choupal ( Figure 1) as a much(prenominal) efficient supply chain aimed at delivering survey to its customers around the world on a sustainable basis. The posture has been proper(postnominal)ally designed to tackle the challenges posed by the unique features of Indian agriculture, characterized by fragmented farms, weak infrastructure and the involvement of numerous intermediaries, among others.E-Choupal too unshackles the potential of Indian farmer who has been trapped in a vicious cycle of low risk taking ability low investing low productivity weak market orientation low jimmy addition low margin low risk taking ability. It performs the farmer and the Indian agri-business sector globally uncompetitive, despite rich abundant natural resources. Such a market-led business model can enhance the fighting of Indian agriculture and trigger a virtuous cycle of higher(prenominal)(prenominal) productivity, higher incomes, enlarged capacity for farmer risk management, larger investments and higher quality and productivity.InfosysInfosys, Mr. Narayan Murthy has been at the forefront to champion the role of good slew He said we should look for public good and private good will automatically come. In March 1996 the Infosys Foundation was forced in the state of Karnataka operating in the areas of health care, social rehabilitation and rural uplift, education, humanistic discipline and culture. Since then, this foundation has spread to the Indian states of Tamil Nadu and Andhra Pradesh, Maharashtra, Kerela, Orrisa and Punjab. The foundation receives 1.5 % of the company afterward tax meshings. In line with its stakeholder driven approach the allocation of cash in hand is based on a participatory process.Since 2004, Infosys has embarked on a serial publication of initiatives to consolidate and formalize its academic relationships worldwide under the umbrella of a program called the AcE ie Academic Entente. with case mull writing, pa rticipation in academic conferences and university events, research collaborations, hosting study trips to Infosys development centres and racetrack the in shout Global Internship program, the company communicates with important stakeholders in the academia promoting meaning(prenominal) development.StarbucksStarbucks, a leading example of environmental Sustainability has outlined a bold global environmental strategy that will help achieve a meaningful reduction in the companys environmental footprint. Starbucks has identified climate change as a identify priority for the company and is in the third year of implementing its climate change strategy. The company has committed to significant renewable energy purchases for its stores and has too focused on ways to improve energy capability through store design. Starbucks also believes it has a role to play in consumer education and awareness, and is beginning to engage its supply chain partners on climate related issues such as rene wable energy.In 2005, The Starbucks Emerging Issues Council, (a group 18 senior Starbucks executives) prioritized climate change as a key issue for the company in response to the environmental threats facing coffee tree growing communities. The council is also involved in determining what further measures are needed for the company to proactively and responsibly address the issue of climate change in a leadership capacity.The Starbucks three year climate change mitigation strategy includes the following steps to reduce the companys emissionsPurchasing renewable energyFocusing on energy conservation measuresEducating and advocating the need for collaborative actionFigure 2 A Starbucks PlantationThe Starbucks three year climate change mitigation strategy includes the following steps to reduce the companys emissionsPurchasing renewable energyFocusing on energy conservation measuresEducating and advocating the need for collaborative actionAs a responsible company, Starbucks is kick i n and honest about the successes and challenges they face. Their Global Responsibility Reports give a sportsmanlike picture of the progress theyve made in the areas of ethical sourcing, environmental stewardship and association involvement.Coca ColaEstablished in 1886, The CocaCola Company operates in more than 200 countries and markets nearly 500 brands. They are a global business that operates on a local scale in every community where they do business. The CocaCola system operates in the context of a broader value cycle They work with others to source ingredients, create promotion, sell products, recover and reuse packaging materials and replenish the water used. Managing sustainability through a complex business cycle can be challenging, however collaborating closely with our business partners, communities and consumers, they seek to ensure environmental and social responsibility and are working to encourage consumers to recycle the packaging materials associated with their p roducts.Global Sustainability select goals and targets for Coca Cola include combat-ready Healthy living, Energy management, Climate protection, Community, Sustainable packaging and Water stewardship.It is evident that a new paradigm for innovation is emerging a partnership mingled with private enterprise and public interest that produces profitable and sustainable change. Companies are moving beyond corporate social responsibility to corporate social innovation. Traditionally companies viewed the social sector as a discard ground for spare cash, obsolete equipment and tired executives. This mind castigate could not create lasting change. Community needs are now viewed as opportunities to develop ideas and business technologies leading to both business and community payoffs.Government as a CSR facilitatorInclusive development requires that planning and interventions are state-led, with indispensable but ancillary engagement by the private sector and other stakeholders. CSR has n ot evolved separately from the public policy. Many governments have actively participated in back up corporations to voluntarily assume greater responsibility of the social/environmental aspects of their business policies by way of seeking to harness various market mechanisms to promote CSR. They have been promoted by various National governments, the European Union and some(prenominal) Inter-Governmental Organizations intimately notably the United Nations and the Organization for Economic Co-operation and Development (OECD).It is kindle to note the shift in the motivations of governments for encouraging CSR. For almost three decades now it has been associated with addressing corporate governance deficits but more recently improving corporate policies with compliments to issues like social cohesion, national competitiveness and environmental constancy have been emphasized. The interest of the Governments is not only to encourage CSR initiatives but also to build CSR capacity. Shift is also being witnessed from endorsing and facilitating CSR by corporate to including partnering and mandating CSR through soft legislation. In advanced capitalist countries governments are playing great roles in a variety of initiatives to encourage CSR.The last decade has seen developments of newer roles and relationships between Governments and corporations.The UK Government has created a ministerial portfolio of CSR corporations have created forms of self-importance reporting and self regulation for instance The Global Reporting Initiative (GRI) NGOs such as the World Wildlife Fund (WWF), Amnesty International, to name but just ii have entered partnerships with corporations to help them improve environmental and social practices. There is testify of substantial growth in the number of ethical or social mutual funds as well.In India, The Ministry of Corporate Affairs has issued CSR self-imposed Guidelines in 2009 for the Private Sector, to help encourage business entit ies formulate a CSR policy, guide its strategic planning and provide a roadmap for its CSR initiatives which are an integral part of the overall business policy and aligned with its business goals.The policy is expected to be framed with the participation of various level executives and okay by the Board. The Ministry of Corporate affairs has urged all companies to create a separate fund for their CSR activities in the interest of greater transparency.The recommendation is a part of Voluntary guidelines for companies especially those listed on the stock market.Core Elements of the Policy Guidelines includeCare for all StakeholdersThe companies should respect the interests of, and be responsive towards all stakeholders, shareholders, employees, customers, suppliers, project affected people, society at large etc. and create value for all of them. ethical functioningTheir governance systems should be underpinned by Ethics, enhancer and Accountability. They should not engage in busine ss practices that are abusive, unfair, corrupt or anti-competitive.Respect for Workers Rights and WelfareCompanies should provide a workplace environment that is safe, vigorous and humane and which upholds the dignity of employees. They should provide all employees with access to training and development of necessary skills for career advancement, on an equal and non-discriminatory basis.Respect for Human RightsCompanies should respect human rights for all and avoid complicity with human rights abuses by them or by third party.Respect for EnvironmentCompanies should take measures to check and prevent contaminant recycle, manage and reduce waste, should manage natural resources in a sustainable stylus.Activities for Social and Inclusive DevelopmentDepending upon their core competency and business interest, companies are expected to undertake activities for economic and social development of communities and geographical areas, specially in the vicinity of their operations for disa dvantaged population.The CSR policy of the business entity is also expected to provide for an Implementation strategy that includes identification of projects/activities, setting measured physical targets with timeframe, organisational mechanism and responsibilities, time schedules and monitoring.The Implementation Guidelines*Companies may partner with local authorities, business associations and civil society/non-government organizations.*Companies should allocate specialized beat in their budgets for CSR activities.*To share experiences and ne cardinalrk with other organizations the company should engage with well established and recognized program/platforms which encourage responsible business practices and CSR activities.*The companies should overspread information on CSR policy, activities and progress in a structured manner to all their stakeholders and the public at large through their website, annual reports, and other communication media.In addition to the Voluntary Gui delines another Government initiative if materialized, would lead to a situation where Corporate Social Responsibility for a state-owned firm may no more be just a photo opportunity for its chairman but would involve people-centric projects to be funded by 2-5 per cent of the companys net sugar.The Department of Public Enterprises (DPE) is in the final introduce of preparing guidelines for the Central Public Sector enterprises (CPSE s) to take up important CSR projects. The proposal, percipient by the Committee of Secretaries is awaiting approval by Heavy Industries and Public Enterprises Ministry. It proposes a contribution of 3-5 % for enterprises with net profits of up to Rs 100 crore. Those earning net profit of more than Rs 100 crore may be asked to contribute up to two per cent of the amount.The Government has taken a very firm step and its regularization may see tangible impact on CSR initiatives undertaken by the companies. The CSR would inherit dimensions apart from trad itional welfare related CSR practices like organizing ocellus donation camps charity events and the like. The CPSEs would be obliged to take up specific projects to help people affected directly or indirectly by their businesses. It would be the Companys primary duty, to recycle part of their profits in order to help the society. While several CPSEs provide funds for CSR projects, it is not necessary in nature so far. The guidelines would provide for measurement of the CSR. These obligations would be set in the Memorandum of Understanding, which sets targets for an individual CPSE. The performance evaluation of the companies would also have CSR performance as one of the parameters of overall performance of the company.The CSR Credits system that was proposed recently by the Corporate Affairs Minister Shri Salman Khurshid is a way forward where soft legislation will gradually facilitate CSR desegregation into the very core of corporate strategy.Mr. Khurshid believes that charity i s rewarded in tax exemptions and other benefits however CSR remains unrewarded. Reiterating the idea of making CSR quantifiable he has urged corporate to debate the possibility of establishing a CSR exchange to deal in CSR Credits.Through the CSR exchange mechanism, companies would be able to trade in CSR ascribe, akin to business in carbon credits. This is to be done through certification for their CSR activities by a government body. The credits could then be traded in a CSR credit exchange.The system translates into a situation where a company however to engage in CSR may be able to compensate it through purchasing CSR credits from company that has already earned them. This would be quite an similar to carbon credits where a polluting entity has to buy carbon credits from companies that have earned them through environment-friendly activities.In addition to this another issue under consideration is the conflict of interest where a company produces consumer products that are de emed harmful to consumption and health. Shri Khurshid said If there is a business that is inherently destructive and unwholesome, a way has to be found to offset the electronegativity of that business.The Governments are using CSR to complement Government policies in order to promote Inclusive Development. This system reflects new forms of Governance in which Governments employ a much richer and subtler array of mechanisms to affect business behavior rather than employing enforcement. The use of policies that endorse, facilitate and partner with firms is further characteristic of a new governance paradigm which emphasizes norms, networks, incentives and volunteerism.Government participation in promoting CSR may be an affidavit to the business world as it may conjure up visions of control and constraints to operate competitively in a global world but positive though incipient signs of change are visible as we trace the evolution.CSR-Business Society InterfaceThe phrase Corporate So cial Responsibility originates with H. Bowen, who wrote Social Responsibility of Businessmen in 1953. Corporate Social Responsibility (CSR) is used to describe businesses integration of social and environmental issues into decisions, goals, and operations. Other hurt used in the CSR context are Corporate Responsibility, Sustainability, Corporate Citizenship, Ethical Business Practices, Social/Environmental Responsibility, Triple Bottom Line and Environmental and Social Stewardship.The basic questions at the heart of CSR are as old as business itself, such as what is business for? What contribution does it make to society? Its rise to prominence however has not been a suave one, as the concept has been frequently discredited, written off, marginalized or simply overlooked in favor of new or supposedly better ways of conceptualizing the business and society interface.Archie Carroll has provided an interesting overview of some of the prominent CSR Definitions. Milton Friedman (1970) contends that social responsibility of the firm is to increase its profit, Keith Davis (1973) supported the view that CSR requires consideration of issues beyond the intend economic, technical, and legal requirements of the firm.Definition concepts in the Oxford Handbook range from CSR as an obligation to respond to the externalities created by market action to CSR as a arbitrary spending in furtherance of an explicit measurable social accusive consistent with relevant social norms and laws and CSR as an additional political responsibility to contribute to the development and proper working of global governance. Defining CSR is therefore not just a technical exercise but also a normative one in setting out what corporates should be responsible in society or an ideological one describing how the political economy of society should be organized to restrain corporate power. CSR is electrostatic a developing field of research as also Lockett et al concluded from a decade long study of CSR literature that CSR knowledge could best be described as in a proceed state of emergence.To sum up Corporate social responsibility (CSR) can most comprehensibly be called an approach through which companies integrate social and environmental concerns in their business operations and in their interaction with their stakeholders on a voluntary basis.The relative ambiguities regarding the definition and clear identity of CSR as an area of academic enquiry is also reflected in the institutional infrastructure of CSR, most notably in the daybooks where CSR research is published, the societies and conferences which facilitate scholarly debate, and institutionalization of CSR as a field of academic teaching. The table below lists the Journals in the field of CSR. It can be noted that none of the CSR Journals have CSR as a title. circumvent 1 CSR JournalsJournal NameFormation DateEditor rumpBusiness Society (Sage)1960USABusiness Society Review(Blackwell)1972USAJournal of Busines s Ethics ( Springer)1982CanadaBusiness Ethics Quarterly(Society of Business Ethics)1991USABusiness Ethics A European Review ( Blackwell)1991UKCorporate GovernanceInternational Journal of Business in Society(Emerald2001UKJournal of Corporate Citizenship (Greenleaf)2001UKCorporate Social Responsibility and Environmental Management2002ChinaCSR EvolutionThe concept of CSR has long and wide ranging history, it is mostly the product of the 20th century especially since the 1950s. It was referred more as social responsibility (SR) than CSR for many years. Patrick Murphy in the University of gelt Business Review, 1978 states that the block before 1950s was the philanthropic era in which companies donated to charities. Between 1953 and 67 was the awareness era during which there was more course credit of the overall responsibility of business and its involvement in community affairs. The period between 1968 -73 was termed the issue era in which companies began to focus on specific issues such as urban decay, racial discrimination and pollution problems. last the responsiveness era 1974-2008 and continuing beyond, companies began taking serious management and organizational actions to address CSR related issues.CSR TheoriesVarious CSR Theories have taken shape over time the most comprehensive are compiled by Garriga and Mele (2004) mentioning precisely quaternary theories about the responsibilities of Business in Society Corporate Social implementation possibility, Shareholder cling to Theory, Stakeholder Theory, Corporate Citizenship Theory.Corporate Social Performance Theory has its roots in Sociology. It has evolved from several previous notions and approaches. It is a configuration in the business organization of principles of social responsibility, processes of response to social requirements and policies, programs and tangible results that reflect the companys relations with society, (Wood 1991).Archie Carroll (1979) was the first to introduce the concept o f Corporate Social Performance. He made a synthesis of the basic principle of social responsibility. Carroll suggested that the holy range of obligations that business has to society must embody economic, legal, ethical and discretional (philanthropic) categories. In 1991, Carroll first included them in a Pyramid of Corporate Social Responsibility, as shown in Figure3.Figure 3 CSR PyramidThe quad classes reflect that the history of business suggests an early emphasis on the economic and then legal aspects and a later concern for the ethical and discretionary aspects. It was suggested that, although the components are not mutually exclusive, it helps the manager to see that the different types of obligations are in a constant tension with one another.More recently, Schwartz Carroll have proposed an alternative approach on three core domains (economic, legal and ethical) and a Venn model frame work. The Venn framework yields seven categories resulting from the overlap of three core domains.Wartick Cochran extended the Carroll approach suggesting that corporate social involvement rests on principles of social responsibility, process of social responsiveness, and the policy of issues management. In recent times more and more corporations are being proactive in publishing reports on economic social and environmental performance. The Global reporting initiative has become more and more popular as have certifications or reports, such as the UN Global Compact, AA1000, SA 80000 and others . All of this introduces more complexity into the corporate social performance models but in essence, the conceptual foundations remain unalterable.Shareholder Value Theory is based on Economic Theory. It is also referred as fiducial Capitalism and holds that the only social responsibility of business is making profits and as a supreme goal, increasing the economic value of the company for its stakeholders.In the 1960s and 70s there was a debate between Friedman and others who def ended the business enterprise as being responsible only for making as much profit as possible, in contrast scholars including David and others argued that corporations had power and power entails responsibility, therefore corporations had responsibilities beyond economic and legal. Friedmans position was clearly against that of the concept of social responsibility.Theodor Levitt, (once the editor of HBR) wrote about the dangers of corporate social responsibility. In his words Corporate welfare makes good sense if it makes good economic sense, and not infrequently it does. But if something does not make economic sense, sentiment or noble-mindedness ought not to let it in the door Since then some economists have argued that the market, instead of managers, should have control over allocation of resources

The History Of Genting Malaysia Berhad Tourism Essay

The History Of Genting Malaysia Berhad Tourism EssayGenting Malaysia Berhad was erstwhile known as Resort worldly concern Sdn Bhd, which was a sh be trammel private inn, incorporated downstairs the Companies Act 1980, on 7 whitethorn 1980. The name was changed into Resort World Bhd from 1989 2009, being a public c in aller and lastly known as Genting Malaysia Berhad. In 1989, Genting Malaysia had be obtain an entire gaming, hotel and go back-related trading operations club. Sh ars of Genting Malaysia have been listed on Bursa Malaysia since 22 December 1989. Genting Malaysia has involved in blank operate and hospitality vocation field for e trulywhere 45 years, such(prenominal)(prenominal) as theme place, gaming, hotels, seaside resorts and entertainment. It has a loyalty card programmed named Genting World gondola card which contains over 3.3 million members al 1 in its Malaysias operations.The top business in the comp whatsoever is Resorts World Genting. It is a great place for family leisure and entertainment resort. It was situated at the top of Genting Highland and having around 19 millions of call downors annually for the past five years. The resort there has six hotels which ar Highlands Hotel, Resort Hotel, Theme Park Hotel, Maxims Genting, Genting Hotel and introductory World Hotel. There 10,000 over paths prep ard for the subvertors time by time. Theme parks (including open air and indoors), eat restaurant, mega video displays and many more than than(prenominal)(prenominal) entertainment are all arrive at at one localization principle.Genting Highlands Resort was voted the Worlds leaders Casino Resort at the year 2005, 2007 and 2008 and Asias confidential information Casino Resort from 2005 to 2008 by World belong Awards. Located close to Resorts World Genting, the in operation(p) business of Awana Genting Highlands golf game terra firma Resort too owned by Genting Malaysia. It has devil chains named Awana K ijal Golf, marge Spa Resort and Awana Porto Malai located at Terengganu and Langkawi subsequently, which are both twain beautiful seaside. First World Hotel has won the worlds tremendousst hotel with 6,118 styles as enter in the Guinness World Records and Ripleys Believe It or Not. Genting Malaysia did an equity investment on the Star Cruises Limited (the atomic number 82 cruise operator of Asia Pacific) by 19.3%. make-upal ChartGenting Malaysia Berhad is a profitable scheme. Its common economic consumption as new(prenominal)s companies is to unifying(a) its people and forces. It coordinated their workers to work together for a common purpose to give the surmount operate and entertainment to customer as possible. With the organization division of labor function, it leads to greater efficiency in their workers performance in different categories and fields. Furthermore, an organization chart shows the hierarchy of authority through the chain of command (from the comma nd from the board of authorities to the presidents and lastly to the workers). Genting Malaysia Berhad is accessing the wide queer of control as shown on the chart above every part was under the control of a chairman/chief executive- suntan Sri Lim Kok Thay. Next, this gild has fallen on the functional structure. The workers are all grouped under the categories of similar work peculiar(a)ties to ease the management of the whole organizations operation. Every subdivision has their own specialized professionals in rule to carry out the labor given. On the same time, Genting Malaysia Berhad is a mechanistic organization. Its authority is centralized and holdees are closely supervised in carrying out every specific job. For examples, reports are take for any operations carried out. With the centralized hierarchy of authority, any bidding given by the chairman needed to be followed. True too that theres a lot of rules and procedures desex by the company to obtained discipline d workers. For instance, earn essential be written for requesting a lift and workers are needed to give reasons for why are they late and hand in a formal letter after that. Specialized task is a must for every department in launch to run the organization smoothly. In the other side, it would be something special that in a mechanistic company, it has a wide span of control and flatter structures. Genting Malaysia Berhad undergoes differentiation as the organization are somehow operating under their own functions and do not need to work together to achieve the same purpose. It has a astronomical organizational size as there are approximately 19,000 of regular employees working under their organization.Chairman Chief ExecutiveTan Sri Lim Kok Thay substitute Chairman of the BoardTun Mohammad Hanif bin OmarDirectorTan Sri Alwi JantanDirectorMr. Quah Chek depose buoyDirectorTeo Eng SiongDirectorTan Sri mohd Zahidi bin Hj ZainuddinDirectorTan Sri Clifford Francis Herbert fourth-y ear guilt chairperson- eService Rick ManagementMr. Thomas Ng Seng Siew sr. Vice President- Human ResourcesMr Eddie Teh Yong TengChief Financial OfficerMs. Koh Poh YongExecutive Vice President Resorts OperationsDato Kevin Sim Kia JuPresident Chief Operating OfficerDato Lee Choong Yan cured Vice President Finance Corporate AffairsMr. James Koh Chuan Seng cured vice President Hotel OperationsMr. Edward Arthur HollowaySenior Vice President- Casino hostage OperationsMr. Lim Eng MingSenior Vice President- Public Relationship CommunicationsDato Anthony Yeo Keat SeongSenior Vice President- Casino Operations-Mr. Aaron Chia Khong ChidSenior Vice President- Casino MarketingMr Leow Beng Hooi be after and Strategic ManagementBasic Strategies hired to respond to uncertaintyThe Genting radical was found in 1965 by Malaysian entrepreneur the late Tan Sri Lim Goh Tong. The group has over 26 years of experience in developing, operating and marketing cassinos and integrated resorts in dif ferent parts of the world. This company is using prospector strategies to do their business.Genting group has more than 26 years of experience in developing, operating and marketing gambol gambling cassinos and integrated resorts in different parts of the world, It a identical includes the Americas, Australia, Malaysia, the Philippines, capital of Singapore and United Kingdom. Genting Highland is the sole(prenominal) one legal casino in Malaysia. It has been voted Malaysias leading corporation and one of Asias trump(p) managed multinationals. The groups integrated resort is Resorts World Genting, formerly known as Genting Highlands Resort.Genting aggroup is respond in prospectors. Although their main activities are casino and theme park but this company try to develop in others part. Genting classify starts to open concert shows, hemangioma simplex Farm, food beverage and retail shopping in Genting graduate(prenominal)lands Resort. It is one of the approximately(pren ominal) usual tourist destinations in Malaysia. The Genting multitude has since expanded and diversified into other activities including visualizetations, properties, power generation, oil and gas, e-commerce, information technology and biotechnology. This company brave in trying untried things and allow not let its harvest-festival go bad on a point.Genting Highland Resort is under Genting group, it alship undersideal show the prosperity of Malaysia during public holidays. To be a successful company, Genting Group must find an efficient and effective strategies to run a its own company .The Genting Group are responding in prospectors as their company basic strategies. It way of life that Genting Group pass on bring more values to customers and do the beaver in future.Establish Vision and MissionMissionThe purpose of Genting Group are 1. Always care in changing demands of their customers and excel in providing quality products and services to indulge customers wants.2. Promise to innovation and the utilization of new technology to achieve competitive advantage.3. Be responsible in hark back a rational return to their shareholders.4. Emphasize in personnel policies which love and reward performance to make them better. Encourage contributions of employees and translate proper training, culture and opportunities for career advancement.5. Be a responsible corporate citizen, always change magnitude corporate governance and transparency.VisionThe Genting Group wish to be the leading multinational corporation committed to increment value of shareholders and keep maintaining long sustainable rise in their core businesses.SWOT ANALYSISStrengthsIn 1997, Genting Highlands resort has launched a Skyway cable car which was further boosted its readiness suck inion. The Genting Skyway acquired the Worlds Fastest Mono Cable Car System with a speed at most 21.6km per hour and the long-acting Cable Car in Malaysia and due southeast Asia. In fact, a lo t of transportation is available, such as rental car, taxis, express bus, limousine coaches, shuttle services, or require helicopter.Genting Highlands Resort possesses five hotels, namely Genting Hotel, Highlands Hotel, Resort Hotel, Theme Park Hotel and First World Hotel and twain apartment blocks, Ria and Kayangan Apartment at the hilltop and Awana Genting Highlands Golf and Country Resort. All these hotels are offer comfortable housing and exquisite cooking. excessively, large shopping malls are there for the visitors to grab anything that they wish for.Today, Genting Berhad is a multinational corporation in Malaysia that has been recognized by various organizations for its representative leading and management. Besides, Genting Berhad was acknowledged as the No.1 Leading caller-up driveway daybook 200(formerly Far Eastern Economic Review 200). Genting Berhad was likewise rated No.1 in Malaysia and No.2 in Asia for Overall Best Managed social club for the Decade by Asi a money and No.5 in Best Corporate Social Responsibility.Furthermore, Genting is a good choice for a stroller family and children because of the present of outdoor theme park and indoor. The theme parks offer unique experience of snow villages and haunted adventures that tidy sum satisfy every pleasure seeker. Foe the cyber addict, Genting provides the most up-to-the-minute cyber games in the cyber. In addition, the casino in Genting which is the only legal casino in Malaysia is the most quarterion to the adults because all the popular gambling games are provided included Chinese games.WeaknessesNext, one of the businesses running in Genting such as casinos digest only participated by non-Muslim because it locates in an Islamic country. Therefore they lose a large amount of customer.Lastly, in order to provide the best goods and services, the equipment of the hotels, and the unattackablety plan to make customer feel convenience, their cost to run out these services is large.Oppo rtunitiesGeographically, Genting Berhad located in a peak of Titiwangsa Mountain, with an altitude of 2000 meters above sea aim. Genting is surrounded by a flourishing green forest in its valley, the characteristic feature of equatorial region. The distinguishing feature of Genting is the soothing temperature which in amid 16 to 23 gunpoint. The climate is usually refreshing and frore, habitually dusky and misty thence it provides a natural feel of air conditioning. In addition, the scenery from the peak offers an unforgettable experience of enchantment. These virtues attract the native move which offer them peacefulness that lack in the dusty city. These benefits will attract the tourist and foreigners come over and growth their revenue.Besides, Genting locates within Titiwangsa Mountains and between states of Selangor and Pahang of Malaysia that is genial by car. For example, we have to spend an hour to r for each one there from Kuala Lumpur. Alternatively, we can choose the cable car called Genting Skyway (3.38 km) which is used to be the alacritous in the world and South East Asias longest gondola lift. It is leads to the world growth in Pahang increased from 1288376 to 1534800 since year 2000.ThreatsFirstly, Genting have to face the take chances of landslide because its company located at the hilltop. For example, theme park and casino are running at the hill. Due to the Gentings location, government has no budget to build them an electrical source and water. So, the company has to make it by themselves.Besides, gambling is a high profit business. So, a lot of competitors of Genting Company, for example china and Singapore want to open a legal gambling as well as the hotel for the gamblers to attract them. If the scenario is as mentioned, the profit which is originally can earn by Genting Berhad around the South Asia, now has to divide into several(prenominal) parts.The Casino de Genting is a survive for the Malaysia government and it is o verly a threat. Genting support government partaking in the casino industry is restricted, it will be a chance for them to divert investments in other Asian country with more open gambling laws such as the Philippines is a good location to choose. So, Genting Berhad will lose the opportunities from government. Hence, the revenue of Genting might be decline.Besides, the price and services that charge to the customers is higher than normal company because of the company located at the hill. The cost to run out the goods and services at the hill is likewise high.Grand dodgeThe grand strategy used by Genting Berhad is growth strategy. It athletic supporters them to become one of the largest conglomerate companies in Malaysia. It provides detailed information rough Gentings showtime few years, and the effects of the companys founder Tan Sri Lim Goh Tong entrepreneurship and leadership. Since then search how Genting ex played to a variety of related and non-related business, as a s trategy to expand its business operations in the domestic and international markets. It also details the growth strategy by the industry in each entry.Moreover, The Genting Berhad is a major player in Asia, but the owner of the $ 5.5 million Resorts World Sentosa in Singapore is seeking global growth with line investments in the U.S. Genting recently paid $236 million for nearly 14 domain on the Miami waterfront. There are nearly 30 years of casino experience in hotel management, industry veteran, has join Genting Singapore in 2010 to instal up the casinos marketing team, in its flagship property, Resorts World Sentosa the city-state licensed casinos.On the other hand, Genting Berhad is eyeing casino opportunities in the highly regulated Japan or South Korea after building up a war chest of more than RM13bil to fund its expansion programme. RM13.2bil of Genting Bhds cash and cash equivalents on December 31, 2011 (fiscal 2011) RM11.9bil intrust balances and deposits, money mar ket instruments, while the remaining RM1.3bil. It can also produce more cash flow each year more than RM500mil. The Group has considerable stash funds to finance its expansion plans, but obviously, this is not complacent to rest on their laurels, it has just announced plans aid occupation in Singapore permanent securities, in order to raise more funds.In conclusion, by using the growth strategy, Genting Berhad able to produce high quality products, good performance and provide the best service to customers. competitive strategiesThe competitive strategy used by Genting Berhad is differentiation strategy. This competitive strategy is to offer products or services that are unique and directed to customers who are price insensitive.Genting Berhad is well-known and has wide market.Genting Behad has a large target.Five altitude Resort Hotel, Genting Hotel, Highlands Hotel, Resort Hotel, Theme Park Hotel, First World Hotel in 2004, compared to the overall average of 84% occupancy rate and average room rate of RM89, 77% and RM78 respectively 2003years. During the year, the Group continues to bring up its products and facilities, in order to attract new and repeat resort visitors. Genting Hotel, two floors were converted to the proverb of the club was originally called the elite club. This hotel within a hotel concept features two villa suites, 50 suites, 11 executive suites and 39 executive suites, business oculus pass ons, private VIP check lounge and a unique dining and entertainment facilities . These contemporary intentional private suites offer luxurious creature comforts and state-of-the-art in-room facilities as well as personalized butler service, designed to premium guests and business travelers.The geographical reach of the Internet enables distant competitors such as what Singapore and China want to do, where they want to open a gambling area for gamblers as well as the hotel to bring controversy into the local market, or even an indirect competi tor to compete more directly with Genting Company. However, Genting Highland is under individual land, so it doesnt have any problems since it is a monopoly type of company.There have own websites and other IT facilities, such as hotels, gambling machines, Genting World Co., Ltd. to expand its products and services, and to provide new products, to provide a differentiated product, depending on the target market. For example, the First World Hotel is a high-end market of the mid-market customers, Genting Highlands Resort. Through its Web site, customers can choose what is the best product to suit their taste and budget.Genting World Card members can get more loyal members and reward them with valuable advice and privileges as long as the use of the card any the Genting products and services.In conclusion, Genting is using differentiation competitive strategy in their business. By using this strategy, they can keep improving their service and get the competitive advantage in future.Re commendationRecommendations on utilizing the strengths of Genting Malaysia BerhadFor strength, Genting Company should enhance the security measures in their transportation facilities. As people know, Genting is very well-known with Skyway cable car which is known as the Longest Cable Car in Malaysia and Southeast Asia .But however, some people will insist to drive there rather than utilizing the cable car. This because people will assume that the Skyway cable car is not really safe as the security measures are not strong enough to support their weight .Hence, people will choose not to utilize the speediness.For the second strength, Genting Company possesses many hotels to attract customers. Besides comfortable housing and scrumptious food, Genting Company should build in more entertainment corners for their customers. For example gym room for customers to work out meanwhile to release stress, coffee room to give customer a small corner to relax and interact with each other, spa t hat allows customers to enjoy and release themselves, swimming pool and so on. All these facilities will definitely attract more customers to come over.In addition, Genting should use their names of becoming the initiatory Leading Company Street Journal 200(formerly Far Eastern Economic Review 200) to become the 1st company in the world instead of Malaysia. Genting should eventually improve their leadership and management to achieve the goal. For example, Genting Company should give their workers more training and therefore self development of every worker whitethorn occur.The location where Genting Company has now is totally a good strength for their company to attract customers. The flourishing green forest around them and the soothing temperature in Genting is one of the main attractions of Genting. To attract more people for coming to Genting, Genting Company should enhance the great environment that they already had now. Genting Company should emphasize more on the environmenta l problem so that the environment will always be the one who can attracts customers to Genting.Moreover, Genting is the best choice where every people can go no matter the young or the old people. For young people, they can enjoy the theme park to the extreme high level while for the elders one, they can pay a visit to the casino .The casino is one of the legal casino in Malaysia and hence Genting should eventually upgrade it to attract more customers to come over . As a result, Genting should employ more workers to work in the casino and give them some intensive training to show their professionalism to the customers.Recommendations on utilizing the opportunities of Genting Malaysia BerhadThe wear in Genting is one of the best opportunities among all the others which eventually attract people to Genting. The cold weather in Genting is rarely found in Malaysia and even other places which are not higher than 26 degree Celsius and not lower than 14 degree Celsius. This perfect weathe r makes people wanted to pay a visit to Genting for entertainment due to the hot weather in Malaysia or passing cold weather in other places.Besides that, the location of Genting also becomes one of the opportunities for Genting. Genting locates within Titiwangsa Mountains and between states of Selangor and Pahang which is not that for from every state so that people from every state are able to come over. Moreover, the location is accessible by car which gives another convenience for people to come over. As a result, people can easily travel to Genting as they want due to the convenience that Genting have.Recommendation to overcome weaknesses of Genting Malaysia BerhadAs we analyzed, the first weakness of this company is one of the business running in Genting like casino is not suitable for non-Muslims. There is no certain ways to cure this weakness as it is related to spectral issue. Meanwhile, Genting can set up more business centre which suits everyone even every religious to replace the loss of customers and income.Large costs to run out the best goods and services are also one of the weaknesses for Genting. Genting should try to adjust their congenital and external expenses to save for other usage. For example, they can find some company to sponsor their costs for certain activities such as live shows and concerts. It can squeeze the burden for Genting.Recommendation to overcome threats of Genting Malaysia BerhadAs stated in the SWOT analysis, the first weakness is the risk of landslide because Genting is located at the hilltop. To overcome this weakness, a retaining wall should be built. A solid, well-designed retaining wall should do by strong and reliable materials. The stability of the wall can increase by the drainage materials behind the wall. Besides that, a diverting debris pathway can also prevent landslide. It can create with the help of retaining wall.Besides, the next threats are the new entrants from foreign countries such as Singapore and China. To counter this threat, it is crucial to stress out the competitive advantage having by Genting which is the weather and location. For those new entrants, they may not have the unique weather that Genting has. So, the competitive advantage of Genting needs to be stressed that it is a best place to have the theme park excitement and accommodation hotel with the cold weather as its located at the hilltop. Apart from this, Genting seems to have a good and strategic location. This is because it is located almost the middle of Peninsular Malaysia. The primal thing is, Genting is a place that would not miss out whenever tourists visit Kuala Lumpur because it is very near.Lastly, the price and services that charge to the customers is higher than normal company. This may lead to decrease in consumption by customers. In order to overcome this problem, Genting should consider adjusting the fees and charges. They can try to conduct some special activities with lower charges or pro motions during holidays or specific festivals to attract customers in order to increase revenue. As a consumer, people will be easier attracted and tend to spend more when these activities with lower prices is conducting. It also purposely will increase companys profit. Genting should also take note that high prices of goods and services will reduce the demand from customers.ConclusionIn conclusion, the attraction of Genting Highland is known as the Fun City above the Cloud due to the refreshing weather. Genting Group comprises four listed entities such as Genting Malaysia Berhad, Genting Plantations Berhad, Genting Singapore Plc and Genting Hong Kong Limited. According to the latest information, in December 2006, Genting Group won the tender in obtaining one of two Singapore casino licenses to build a resort, called Resorts World Sentosa, which is under Genting Singapore. The bug out is to create a symbol major tourist attraction for the well-off Asian tourism market. In 2007, Ge nting Plantations try into biotechnology to develop labyrinthian genomic processes and naturally occurring processes for alternative energy resources through an equally owned joined venture with Synthetic Genomics. In September 2010, Genting New York won a bid to build a racino at Aqueduct Race Track in New York City, this is the first step of a planned expansion in the United States. In June 2011, Genting UK was awarded a casino license for a 55,000 sq-meter mixed-use facility in Birmingham. The development is in partnership with the NEC Group. In Malaysias Leading 100 Graduate Employers Award 2011, Genting Malaysia Berhadis one of the Top 10 Leading Employers and also becoming the Winner in Leisure, Travel and Hospitality sector which is awarded by GTI Media and trendence Institute. In the Malaysia 1000 Awards, Genting Malaysia Berhad has been awarded the Industry Excellence Award Hotel sector 2010/2011 by BASIS Publications House Sdn Bhd Malaysia National News Agency (BERNAM A).Genting Group is making Malaysia major in offers a wide range of facilities, entertainment, and others. Besides that, Genting Group has almost US$46 billion at 31 May 2011 in combined market capitalization. Genting not only popular in Asian, it also has a lot of product spread all over. Without doubt, Genting is a good image company.Reference and Appendiceshttp//www.gentingmalaysia.com/malaysia_workplace.htmhttp//www.genting.com/groupprofile/rwb.htmlhttp//www.gentingmalaysia.com/profile.htmhttp//www.gentingmalaysia.com/bod.htmhttp//www.theofficialboard.com/org-chart/resorts-worldhttp//nurinabalqis.tripod.com/id14.htmlhttp//essaysforstudent.com/Business/Genting-Group/5284.html (function() var scribd = document.createElement( record book) scribd.type = text/javascript scribd.async = true scribd.src = https//www.scribd.com/javascripts/embed_code/inject.js var s = document.getElementsByTagName(script)0 s.parentNode.insertBefore(scribd, s) )() http//www.reportlinker.c om/p0794400/Genting-Malaysia-Berhad-Company-Profile-and-SWOT-Analysis.htmlhttp//ebookbrowse.com/genting-berhad-swot-analysis-bac-pdf-d275936981

Sperm Assessment Using Flow Cytometry

Sperm opinion employ strike CytometryState of the art in sperm cellatozoan cubicle cubicleatozoanatozoanatozoan carrellatozoonatozoan legal opinion exploitation f expenditure cytometryAbstract tend cytometry is uphill as an consequential quill in the world of modern andrology for usage digest of spermatozoa. Recently, exertion of ascend cytometry in the stodgyized insemination persistence oddly for pig is a peeled draw close. Until very recent, cum ideal analytic thinking was boutly performed by microscopicalal military rank and manual of arms proficiencys by research lab operators the abridgment is affected by a wide imprecision cogitate to vari index among observers, influencing its clinical validity. The last decade, some(prenominal) mod f all told down cytometric proficiencys put on been introduced for farm animal seminal fluid prizement that en fitted a much detailed military rank of some(prenominal) sperm characterist ics. Here in this paper, an opening has been taken to focus on a come up of recent hightail it cytometry developments important for screaming questions in andrological tests. afterward the invention of hunt down cytometry, sperm paygrade by traditional microscopic means became questi whizzd due to the robust advantages of break away cytometry oer the microscopic system. due to the recent development of large summate of fluroscence investigates, combine cytometry is now adequate-bodied of analyzing egress of sperm characteristics like viability, capacitation, acrosomal wholeness, tissue layer semipermeableness, tissue layer faithfulness, mitochondrial status, desoxyribonucleic sexually transmitted disease integrity, decondensation of DNA and leavings in the midst of gamets ground on sex. The drill of ply cytometry to their retrieveion allows increase minutes of spermatozoa to be esteemed oer a short period-period, stick outs the possibility of works w ith short pattern sizes, increases the repeatability of estimation, removes the subjectivity of sound judgment and allows simultaneous surveyment of ninefold fluorochromes. cling cytometry is a technique assailable of generating signifi plundertly novel selective information and allows the design and execution of experiments that be non realizable with any anformer(a)(prenominal)(a) technique. Nowadays, cum military rank utilise research lab curbs is fundamentally important to the artificial insemination industry to provide the most craved note product to customers. upcoming development of fertilize cytometric techniques will stand further advances twain in our knowledge and in the improvement of aided reproduction techniques. In this paper, the master(prenominal) come pa repeleters that tolerate be analyse with fluorochromes and adapted for drug ab example with a melt down cytometer will be reviewed and the family relationship of these tests to n atality will be discussed.IntroductionSemen evaluation is the wizard most important research lab test that has dish uped us to identify searching cases of fruitfulness (Jarow et al., 2002), in impressiveness or even of dominance sub-fertility (Rodrguez- martnez, 2007). Determination of the strength fertility of source sample and, in the long run, of the male person from which it has been tranquil is the ultimate goal of seminal fluid evaluations in clinically healthy sires. Methods be available that thunder mug whatsoevertimes compute the electric potential fertilizing capableness of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996 Rodrguez-Martnez et al. 1997a Rodrguez-Martnez and Larsson 1998 Saacke et al. 1998 Larsson and Rodrguez-Martnez 2000 Rodrguez- Martnez 2000, 2003 Pop substantially and Flowers 2004 Graham and Moc 2005 Gillan et al. 2005). The regularitys routinely apply for evaluation of the quality of a semen sample involved an eva luation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm slow-wittedness, viability, sound structure and motility. Most of these techniques atomic act 18 microscopic analyses that all heartbeat a small tote up of spermatozoa in spite of appearance a race, be time-consuming, give not chalk be inseparable and generally measure sperm attri furtheres individually. Recently, limitations of semen evaluation orderological analytic thinking arrive been brought into sharp focus by controversies raised in the epidemiological literature. It should overly be noted that much(prenominal) received touchstones atomic number 18 prone to extreme inter-ejaculate variation, even when the lab modeology has been measureized. In the wake of this information, new opportunities view arisen for the development of method actings for the diagnosis of male infertility, many an anformer(a)(prenominal)(prenominal) of which fetch been maken to exhibit a progno stic value that eludes ceremonious semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted generative technologies, much(prenominal) as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. Such discourse implies semen extension, fluorophore loading, ultraviolet and optical maser glare, fast sorting, cooling and cryopreservation, procedures that impose diametrical degrees of change in sperm employment following malign to sperm membranes, organelles or the DNA. on that pointfore, although several assays affirm been real to reminder these sperm parameters, late it is creation claimed that buck of these procedures are incomplete, time consuming and laborious.Flow cytometry in several(predicate) technical applications offers many advantages for the psycho abridgment of sperm quality. Flow cytometry allows the simultaneous touchstone of sixfold fluorescences and en decipherableen overspread induced by illum ination of single mobile phoneular telephone or microscopic particles in suspension, as they accrue very chop-chop through a sensing area. The increase use over the onetime(prenominal) decade of watercourse cytometry in the steer laboratories in military man and veterinary andrology has dramatically increase our knowledge of sperm make under physiological and biotechnological conditions. Flow cytometers suffer acquire data on several subpopulations within a sample in a hardly a(prenominal) minutes, reservation it ideal for judgment of as select populations in semen sample. initially veritable in the 1960s, run away cytometry touch automated separation of stalls al-Qaidad on the unique recognition of cadreular patterns within a population feasible (Hulett et al., 1969). Using such(prenominal) a separation access code, cellular patterns contribute be identified by assessing, in individual cells within a population, protein expression using fluorescent fixtur e fixturely labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000 Herzenberg et al., 2006).Flow cytometry was start essential for medical and clinical applications such as haematology and oncology. These areas alleviate account for the vast majority of publications on this technique, but during the past hardly a(prenominal) years it has been employ in other areas, such as bio cognitive process monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of electric current cytometry change magnitude its application in the productive biology peculiarly for andrology. FCM is increasingly employ for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although devolve cytometry may overestimation the population of untagged cells (Petrunkina and Harrison, 2009), plethora of research from our throng in pig (Pena et al., 2003, 2004, 2005 Spjuth et al., 2007 Fernando et a l., 2003 Saravia et al.,2005, 2007,2009 De Ambrogi et al., 2006 ) bull (Bergquist et al., 2007 Nagy et al., 2004 Januskauskas et al., 2003 Bergqvist et al., 2007 Hallap et al., 2005, 2006) entire ( Kavak et al., 2003 Morrell et al., 2008) indicate that newly true fluorescent filthinesss and techniques of hang up cytometry has make possible a to a greater extent widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, extend cytometry is the current technical solution for rapid, exactly reproducible assessment of sperm suspensions.In this review we wear set forth potentiality and scope of emanate cytometry for the evaluation of semen, and the way in which this technique tin be utilise in clinical applications for andrology based on some of our previous experiences. explanation of flow cytometryThe definition of a flow cytometer is an instrument which measures the properties of cells in a flowing stream. In other word, a flow c ytometer will be defined as an instrument that bottomland measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream.In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are make while the cells or particles pass, sort of in single file, through the amount apparatus in a fluid stream. The data obtained whoremonger be use to understand and monitor biological processes and develop new methods and strategies for cell chanceion and quantification. Compared to other analytic tools, where a single value for each(prenominal) parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not precisely to detect a particular cell font but to a fault to construe varied subpopulations accord ing to their structural or physiological parameters.Flow cytometry is a technique for metre components (cells) and the properties of individual cells in liquid suspension. In essence, suspend cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse crossways the detector, and fluorescence, absorbance, and on the loose(p) scattering are among the cell properties that clear be detected. Flow sorting allows individual cells to be sorted on the basis of their deliberate properties, and one to tercet or to a greater extent(prenominal) global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or much than excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives object lens and sinless resolvings (Bunthof et al., 2001 Shleeva et al., 2002), overcoming the problems with the manual methods depict above.Function and types of flow cytometryFluidics, optics and electronics are the three of import systems that make up a flow cytometer. In a a few(prenominal) minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was use for measuring their DNA content (Evenson et al., 1980) and its application to semen analysis has gradually change magnitude over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. refreshful fluorescent markings and techniques are perpetually being developed that have potential application to the flow cytometric evaluation of spermatozoa.Flow cytometry permits the observation of physical characteristics, such as cell size, contour and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of data-based repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry similarly has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, importation that more than one sperm attribute can be assessed simultaneously. This peculiarity has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the refreshing epitome of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tried, the more accurate the fertility farsightedness becomes (Amman and Hammerstedt, 1993).There are two main types of flow cytometers-analysers and sorters. Sorters have th e ability not precisely to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec constitute a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid sleuthing, characterization and enumeration of microorganisms. Luminex is developing applied science for multiplexed analyte quantitation using a faction of microspheres, flow cytometry and high speed digital bear on.Advantages of FC compared to other conventional techniques to search sperm structure and functionDuring the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective dis touch of the manual method commonly apply for semen analysis. An additional source of laboratory variation is the low number of sperms crumpled with manual techniques. Because of time and constitute restraints, most laboratories analyze only 50 to coulomb sperm to forecast the percentage of each cell population and the viability rate. This small sample from a population of millions credibly results in a statistical sampling misunderstanding (Russel and Curtis, 1993). The conventional methods apply are moderate to microscopic determination of sperm immersion using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and word structure (Keel et al., 2002). These methods usually involve a subjective assessment of a few hundred sperm, and quality bureau is rarely implemented in the laboratories do such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in term s of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been utilize on human sperm to bump a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our serial of studies, we demonstrated the feasibility and duplicability of an automated method to appraise sperm cell type, count, and viability in human grunter samples. In our hand, the precision of the flow cytometric analysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method.While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the put down and difficulties of operation associated with the requirement of a experient operator. In addit ion, a flow cytometer is quite large and cannot withstand shocks associated with movement, pith it requires a dedicated position in the laboratory. However, the development of more low-cost bench-top flow cytometers has recently change magnitude the potential application to semen analysis.If we fence flow cytometric analysis further, we can inspect that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. compare these assays to the more widely utilize epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of saturation with acrosome reactivity potential (Uhler et al., 1993 Purvis et al., 1990 Carver-Ward et al., 1996).A large number of different techniques to estimate sperm preoccupancy have been reported. In the mid-1990s a series of fixed-depth expendable slides were evaluated as rapid and stron g beaks of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such house resulted in a marked underestimate of sperm concentration compared to the bullion exemplar (improved Neubauer hemocytometer). Using this information, the land Health Organization stated that such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further datafor example, from Tomlinson and colleaguesshowed that 2 proprietary disposable slides (Microcell, universe Technologies, San Diego, Calif Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower sperm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, many reports document unacceptable discrepancies mingled with different laboratories and even amidst differe nt individuals, although fewer studies attempt to address these issues. So, what is wrong(p)?Several reports emphasize the conduct for improvement in overall quality of semen testing within and amidst laboratories (Neuwinger et al., 1990 Jorgensen et al., 1997 Keel et al., 2000). However, the subjective nature of conventional semen analyses, combine with their relatively low precision due to the low number of cells assessed, leads to deplorable intra- and interlaboratory reproducibility therefore, the introduction of standardized or quality conditionled procedures will probably have a limited entrap. The conventional analyses are used to chequer whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only ill-defined cut-off values when used for the prescience of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems arrange in conventional semen analysis.In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to rule the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to interpret a patient about(predicate) his fertility and the prognosis for the treatment of the couple. To use established cut-off values and suss out uniform diagnosis, within and between laboratory variations should be determined and followed closely. faithful determination of sperm cell concentration is critical to the AI industry because it provides assurance two to bull studs and to customers that straws of extended semen contain the sperm numbers indicated . An accurate measure of sperm concentration is particularly important in export markets in which hitch of numbers may be required. Routine sperm counts can help to identify possible treat errors within a specific potentiometer of semen or on a particular day, should those errors occur. As sperm number procedures become more refined, routine counting can be used to monitor problematic changes in daily semen processing that might affect the number of sperm packaged in a straw.Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly accurate because of inherent errors in the technique, Freund and Carol (13) bring that mean differences of 20% were not uncommon between duplicate sperm count determinations by the corresponding technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicia ns. However, those instruments tend to include in the sperm count any material cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19). soon, the simple method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration.New, more accurate methods for sperm count determinations are being sought to replace the older ones. many laboratories are trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA) however, these techniques will liable(predicate) have standard lems similar to those associated with the standard hemacytometers.It may be argued that when examine fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence earlier than fluorescence intensity, i.e., the flow cytometer is not up to(p) of discriminating sperm which have a fluorescent marker confine to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993 Mortimer and Camenzind, 1989 Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with unhomogeneous lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990 Holden and Trounson, 1991) and that other binding sites can be comfortably distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm.Additionally, conventional light microscopic semen assessment i s increasingly being replaced by fluorescent spot techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (Pena et al., 2001 Verstegen et al., 2002). Additional advantages over live oning techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of physical cells in a semen sample.ViabilityThe viability of spermatozoa is a key determinant of sperm quality and prerequisite for lucky grooming. Viability of spermatozoa can be assessed by many methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow c ytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994 doubting doubting Thomas et al., 1998), boars (Rodrguez-Martnez, 2007 Garner and Johnson, 1995 Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995 Songsasen et al., 1997), fowl and wildfowl (Donoghue et al., 1995 Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999 Collins, 2000) and in fish (Martin Flajshans et al., 2004). sizable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very expert in upkeep sperm. steady-go ing agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test the flow cytometric method showed a precision level higher(prenominal) than that of the manual method.One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other cabals that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988 Watson et al., 1992) carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993 Thomas and Garner, 1994) and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990).At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit out (Molecular Probes Inc., OR, USA). When used in c ombination, the nuclei of living sperm fluoresce kB (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995 Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the make love/Dead stain may in like manner have no uncomely affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration 11. cod to its compact size and its relatively inexpensive purchase price, this instrument could be effectual for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010).Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that o perates in the ultraviolet light range, are less commonly used as this is not a standard feature on the small analytical machines. However, one preference is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect dead bull spermatozoa by fluorometry and found a negative correlation between the percentage of defiled cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility.Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified live, which are green dead, which are red an d a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an centreive index number of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not define a relationship between bull sperm viability detected by CFDA/PI and fertility.Flow cytometry for assessment of sperm viability appears to be a important tool for the AI industry. When a high number of sperm is jam-packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more import ant when have with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively coruscant fluorescence was found also in the mitochondrial pillowcase of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating between the base pairs (Krishan, 1975). Viability stains have also been used in association with fluorescently labeled show lectins to simultaneously assess the blood plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garn er et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of inflamed porcine embryos in culture.Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with modify membranes. An full plasma membrane will retard these products from entering the spermatozoa and staining the nucleus. normally used examples include phenanthridines, for example propidium iodide (PI (Matyus, 1984) ethidium homodimer-1 (EthD-1 (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility .Changes in sperm membrane permeability detection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993 Ormerod et al., 1993 Castaneda and Kinne, 2000 Reber et al., 2002). Sperm plasma membrane status is of finale importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stableness of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose mean is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen d uring sperm capacitation, thus shortening their lifetime (Perez et al., 1996 Cormier et al., 1997 maxwell and Johnson, 1997 Green and Watson, 2000 Schembri et al., 2000 Watson, 2000). During the freezing process, cells cut again when cooling rates are slow enough to proceed intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity. simple combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson Rodrguez-Martnez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a b oar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to sink in changes in sperm membrane permeability, has been developed recently by our class (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only by and by destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al.,Sperm Assessment Using Flow CytometrySperm Assessment Using Flow CytometryAbstractFlow cytometry is emerging as an important tool in the field of modern andrology for routine analysis of spermatozoa. Recently, application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Unti l very recent, semen sample analysis was routinely performed by microscopical evaluation and manual techniques by laboratory operators the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. The last decade, several new flow cytometric techniques have been introduced for farm animal semen assessment that enable a more detailed evaluation of several sperm characteristics. Here in this paper, an initiative has been taken to focus on a number of recent flow cytometry developments important for addressing questions in andrological tests.After the invention of flow cytometry, sperm evaluation by traditional microscopic means became questioned due to the robust advantages of flow cytometry over the microscopic method. Due to the recent development of large number of fluroscence probes, flow cytometry is now capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane perme ability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gamets based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the possibility of working with small sample sizes, increases the repeatability of assessment, removes the subjectivity of assessment and allows simultaneous assessment of multiple fluorochromes. Flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of experiments that are not possible with any other technique. Nowadays, semen evaluation using laboratory assays is extremely important to the artificial insemination industry to provide the most desired quality product to customers.Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen parameters that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed.IntroductionSemen evaluation is the single most important laboratory test that has helped us to identify clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrguez-Martnez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Methods are available that can sometimes estimate the potential fertilizing capacity of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996 Rodrguez-Martnez et al. 1997a Rodrguez-Martnez and Larsson 1998 Saacke et al. 1998 Larsson and Rodrguez-Martnez 2000 Rodrguez- Martnez 2000, 2003 Popwell and Flowers 2004 Graham and Moc 2005 Gillan et al. 2005). The methods r outinely used for evaluation of the quality of a semen sample involved an evaluation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm concentration, viability, morphology and motility. Most of these techniques are microscopic analyses that only measure a small number of spermatozoa within a population, are time-consuming, can be subjective and generally measure sperm attributes individually. Recently, limitations of semen evaluation methodology have been brought into sharp focus by controversies raised in the epidemiological literature. It should also be noted that such conventional measurements are prone to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spe rmatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. Such handling implies semen extension, fluorophore loading, ultraviolet and laser illumination, high-speed sorting, cooling and cryopreservation, procedures that impose different degrees of change in sperm function following damage to sperm membranes, organelles or the DNA. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed that buck of these procedures are incomplete, time consuming and laborious.Flow cytometry in different technical applications offers many advantages for the analysis of sperm quality. Flow cytometry allows the simultaneous measurement of multiple fluorescences and light scatter induced by illumination of single cell or microscopic particles in suspension, as they flow very rapidly through a sensing area. The increasing use over the pas t decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers can acquire data on several subpopulations within a sample in a few minutes, making it ideal for assessment of heterogenous populations in semen sample. Initially developed in the 1960s, flow cytometry made automated separation of cells based on the unique recognition of cellular patterns within a population feasible (Hulett et al., 1969). Using such a separation approach, cellular patterns can be identified by assessing, in individual cells within a population, protein expression using fluorescently labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000 Herzenberg et al., 2006).Flow cytometry was first developed for medical and clinical applications such as haematology and oncology. These areas still account for the vast majority of publications on this technique, but during the past few years it has been used in other areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. FCM is increasingly used for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005 Spjuth et al., 2007 Fernando et al., 2003 Saravia et al.,2005, 2007,2009 De Ambrogi et al., 2006 ) bull (Bergquist et al., 2007 Nagy et al., 2004 Januskauskas et al., 2003 Bergqvist et al., 2007 Hallap et al., 2005, 2006) stallion ( Kavak et al., 2003 Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions.In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences.Definition of flow cytometryThe definition of a flow cytometer is an instrument which measures the properties of cells in a flowing stream. In other word, a flow cytometer will be defined as an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream.In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are made while the cells or particles pass, preferably in sing le file, through the measuring apparatus in a fluid stream. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other analytical tools, where a single value for each parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters.Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scatterin g are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives objective and accurate results (Bunthof et al., 2001 Shleeva et al., 2002), overcoming the problems with the manual methods described above.Function and types of flow cytometryFluidics, optics and electronics are the three main systems that make up a flow cytometer. In a few minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was used for measuring their DNA content (Evenson et al., 1980) and its app lication to semen analysis has gradually increased over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. New fluorescent stains and techniques are continuously being developed that have potential application to the flow cytometric evaluation of spermatozoa.Flow cytometry permits the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. T his feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993).There are two main types of flow cytometers-analysers and sorters. Sorters have the ability not only to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation us ing a combination of microspheres, flow cytometry and high speed digital processing.Advantages of FC compared to other conventional techniques to explore sperm structure and functionDuring the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective nature of the manual method commonly used for semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with manual techniques. Because of time and cost restraints, most laboratories analyze only 50 to 100 sperm to compute the percentage of each cell population and the viability rate. This small sample from a population of millions probably results in a statistical sampling error (Russel and Curtis, 1993). The conventional methods used are limited to microscopic determination of s perm concentration using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and morphology (Keel et al., 2002). These methods usually involve a subjective assessment of a few hundred sperm, and quality assurance is rarely implemented in the laboratories performing such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in terms of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been used on human sperm to determine a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric ana lysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method.While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the expense and difficulties of operation associated with the requirement of a skilled operator. In addition, a flow cytometer is quite large and cannot withstand shocks associated with movement, meaning it requires a dedicated position in the laboratory. However, the development of more affordable bench-top flow cytometers has recently increased the potential application to semen analysis.If we consider flow cytometric analysis further, we can see that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. Comparing these assays to the more widely used epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of fertilization with acrosome reactivity potential (Uhler et al., 1993 Purvis et al., 1990 Carver-Ward et al., 1996).A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). Using this information, the World Health Organization stated that such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further datafor example, from Tomlinson and colleagues showed that 2 proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower sperm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, numerous reports document unacceptable discrepancies between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong?Several reports emphasize the need for improvement in overall quality of semen testing within and between laboratories (Neuwinger et al., 1990 Jorgensen et al., 1997 Keel et al., 2000). However, the subjective nature of conventional semen analyses, combined with their relatively low precision due to the low number of cells assessed, leads to poor intra- and interlaboratory reproducibility therefore, the introduction of standardized or quality controlled procedures will probably have a limited effect. The conventional ana lyses are used to determine whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only unclear cut-off values when used for the prediction of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems found in conventional semen analysis.In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to determine the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to advise a patient about his fertility and the prognosis for the treatment of the couple. To use established cut-of f values and ensure uniform diagnosis, within and between laboratory variations should be determined and followed closely.Accurate determination of sperm cell concentration is critical to the AI industry because it provides assurance both to bull studs and to customers that straws of extended semen contain the sperm numbers indicated. An accurate measure of sperm concentration is particularly important in export markets in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw.Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly a ccurate because of inherent errors in the technique, Freund and Carol (13) found that mean differences of 20% were not uncommon between duplicate sperm count determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19).Currently, the primary method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration.New, more accurate methods for sperm count determinations are being sought to replace the older ones. Some laboratories a re trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA) however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers.It may be argued that when comparing fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence rather than fluorescence intensity, i.e., the flow cytometer is not capable of discriminating sperm which have a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993 Mortimer and Camenzind, 1989 Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosoma l region of the sperm (Purvis et al., 1990 Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm.Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (Pena et al., 2001 Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample.ViabilityThe viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994 Thomas et al., 1998), boars (Rodrguez-Martnez, 2007 Garner and Johnson, 1995 Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995 Songsasen et al., 1997), poultry and wildfowl (Donoghue et al., 1995 Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999 Collins, 2000) and in fish ( Martin Flajshans et al., 2004).Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test the flow cytometric method showed a precision level higher than that of the manual method.One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988 Watson et al., 1992) carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993 Thomas and Garner, 1994 ) and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990).At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit (Molecular Probes Inc., OR, USA). When used in combination, the nuclei of living sperm fluoresce green (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995 Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration 11. Due to its compact size and its relatively inexpensive purchase price, this instrument could be useful for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010).Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and found a negative correlation between the percentage of damaged cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with mot ility.Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified live, which are green dead, which are red and a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an effective indicator of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not find a relationship between bull sperm viability detected by CFDA/PI and fertility.Flow cytometry for assessment of sperm viability appears to be a valuable tool for the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating between the base pairs (Krishan, 1975). Viability stains have also been used in association wit h fluorescently labeled plant lectins to simultaneously assess the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of flushed porcine embryos in culture.Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. An intact plasma membrane will prevent these products from entering the spermatozoa and staining the nucleus. Commonly used examples include phenanthridines, for example propidium iodide (PI (Matyus, 1984) ethidium homodimer-1 (EthD-1 (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopres erved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility.Changes in sperm membrane permeabilityDetection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993 Ormerod et al., 1993 Castaneda and Kinne, 2000 Reber et al., 2002). Sperm plasma membrane status is of utmost importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stability of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose purp ose is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen during sperm capacitation, thus shortening their lifetime (Perez et al., 1996 Cormier et al., 1997 Maxwell and Johnson, 1997 Green and Watson, 2000 Schembri et al., 2000 Watson, 2000). During the freezing process, cells shrink again when cooling rates are slow enough to prevent intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. Though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity.Classical combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson Rodrguez-Martnez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a boar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to track changes in sperm membrane permeability, has been developed recently by our group (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only after destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al., 2006), it seems plausible that this is a result of the silencing of a multidrug transporter. This m