Advances in flow cytometry for sperm sexing.

This review presents the key technological developments that have been implemented in the 20 years since the first reports of successful measurement, sorting, insemination and live births using flow cytometry as a proven physical sperm separation technique. Since the first reports of sexed sperm, flow technology efforts have been largely focused on improving sample throughput by increasing the rate at which sperm are introduced to the sorter, and on improving measurement resolution, which has increased the proportion of cells that can be reliably measured and sorted. Today, routine high-purity sorting of X- or Y-chromosome-bearing sperm can be achieved at rates up to 8000 s(-1) for an input rate of 40,000 X- and Y-sperms(-1). With current protocols, straws of sex-sorted sperm intended for use in artificial insemination contain approximately 2 x 10(6)sperm. The sort rate of 8000 sperms(-1) mentioned above corresponds to a production capacity of approximately 14 straws of each sex per hour per instrument.

[1]  L. Johnson,et al.  Sex preselection in rabbits: live births from X and Y sperm separated by DNA and cell sorting. , 1989, Biology of reproduction.

[2]  D. Garner.  Flow cytometric sexing of mammalian sperm. , 2006, Theriogenology.

[3]  J. Gray,et al.  Measurement of mammalian sperm deoxyribonucleic acid by flow cytometry. Problems and approaches. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[4]  D. Houck,et al.  Slit-scan flow cytometry for consistent high resolution DNA analysis of X- and Y-chromosome bearing sperm. , 1996, Cytometry.

[5]  W. Rens,et al.  The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI. , 1999, Journal of animal science.

[6]  D. Arn,et al.  266 Birth of canine offspring following insemination of a bitch with flow-sorted spermatozoa , 2008 .

[7]  D. Pinkel,et al.  Modification of a laser-based flow cytometer for high-resolution DNA analysis of mammalian spermatozoa. , 1986, Cytometry.

[8]  J. Morrell,et al.  Offspring from inseminations with mammalian sperm stained with Hoechst 33342, either with or without flow cytometry. , 1989, Mutation research.

[9]  G. Seidel,et al.  Past, present and future perspectives on sexing sperm , 2003 .

[10]  K. M. Evans,et al.  Birth of lambs of a pre-determined sex after in vitro production of embryos using frozen-thawed sex-sorted and re-frozen-thawed ram spermatozoa. , 2004, Reproduction.

[11]  G. Seidel,et al.  Insemination of mares with low numbers of either unsexed or sexed spermatozoa. , 2000, Theriogenology.

[12]  M. Fulwyler.  Hydrodynamic orientation of cells. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[13]  L. Johnson,et al.  Sex preselection: high-speed flow cytometric sorting of X and Y sperm for maximum efficiency. , 1999, Theriogenology.

[14]  G. Seidel.  Overview of sexing sperm. , 2007, Theriogenology.

[15]  L. Fontanesi,et al.  Current status of sexing mammalian sperm by sperm sorting. , 1999 .

[16]  K. Leslie,et al.  Exploring the impact of sexed semen on the structure of the dairy industry. , 2008, Journal of dairy science.

[17]  T. Robeck,et al.  Development of sperm sexing and associated assisted reproductive technology for sex preselection of captive bottlenose dolphins (Tursiops truncatus). , 2006, Reproduction, fertility, and development.

[18]  E. V. van Munster.  Interferometry in flow to sort unstained X- and Y-chromosome-bearing bull spermatozoa. , 2002, Cytometry.

[19]  G van den Engh,et al.  Parallel processing data acquisition system for multilaser flow cytometry and cell sorting. , 1989, Cytometry.

[20]  G. Evans,et al.  Integration of sperm sexing technology into the ART toolbox. , 2004, Animal reproduction science.

[21]  D. Pinkel,et al.  High resolution DNA content measurements of mammalian sperm. , 1982, Cytometry.

[22]  G. Seidel,et al.  History of commercializing sexed semen for cattle. , 2008, Theriogenology.

[23]  G. Seidel.  Economics of selecting for sex: the most important genetic trait. , 2003, Theriogenology.

[24]  E. Munster.  Interferometry in flow to sort unstained X- and Y-chromosome-bearing bull spermatozoa. , 2002 .

[25]  C. Polge,et al.  Production of bovine calves following separation of X- and Y-chromosome bearing sperm and in vitro fertilisation , 1993, Veterinary Record.

[26]  W. Rens,et al.  Improved flow cytometric sorting of X‐ and Y‐chromosome bearing sperm: Substantial increase in yield of sexed semen , 1999 .

[27]  G. Seidel,et al.  Current status of sexing mammalian spermatozoa. , 2002, Reproduction.

[28]  J. Gray,et al.  Flow microfluorometric analysis of sperm DNA content: Effect of cell shape on the fluorescence distribution , 1976, Journal of cellular physiology.

[29]  D. Pinkel,et al.  Sex preselection in mammals? Separation of sperm bearing Y and "O" chromosomes in the vole Microtus oregoni. , 1982, Science.

[30]  T. Lindmo,et al.  Flow cytometry and sorting , 1979 .

[31]  W. Rens,et al.  A novel nozzle for more efficient sperm orientation to improve sorting efficiency of X and Y chromosome-bearing sperm. , 1998, Cytometry.

[32]  K. M. Evans,et al.  Flow Cytometric Sorting of Fresh and Frozen‐Thawed Spermatozoa in the Western Lowland Gorilla (Gorilla gorilla gorilla) , 2005, American journal of primatology.