Help with determination of sex

M. William Lensch Lensch at chop.edu
Tue Feb 1 09:03:37 EST 1994


In article <2ia98nINNbov at newsman.csu.murdoch.edu.au>,
cummins at possum.murdoch.edu.au (Jim Cummins) wrote:
> 
> 
> Patrick Lamastus writes:
> 
> >t his technique is based upon the different densities of sperm (or other cells) and uses centrifugation to seperate cell type (those w/X and those w/Y). This is termed flow cytometry.
> 
> Wrong, sorry. There have been a number of attempts to separate X and Y
> sperm by differential centrifugation.  None have been repeatable.
> 
> Flow cytometry relies on separation of sperm based on fluorescence
> staining.  This can be a specific in situ hybridisation stain (eg for a
> region of the Y or X chromosome) or a generalised DNA stain that gives
> information about the total DNA content.  Theoretically  X and Y sperm
> differ in DNA content by about 2% but as this is less than the
> within-gender variation you can see that there are big problems in
> successful sorting.   The following refernces may help
> Auger J, Leonce S, Jouannet P, Ronot X (1993): Flow cytometric sorting
> of living, highly motile human spermatozoa based on evaluation of their
> mitochondrial activity. J Histochem Cytochem 41:1247-51.
> Cran DG, Johnson LA, Miller NG, Cochrane D, Polge C (1993): Production
> of bovine calves following separation of X- and Y-chromosome    
> bearing sperm and in vitro fertilisation. Vet Rec 132:40-1.
> Dresser DW, Atkins CJ, Pinder A, Morrell JM (1993): Analyses of DNA
> content of living spermatozoa using flow cytometry techniques. J Reprod
> Fertil 98:357-365.
> Johnson LA, Clarke RN (1988): Flow sorting of X and Y
> chromosome-bearing mammalian sperm: activation and pronuclear
> development of sorted bull, boar and ram sperm microinjected into
> hamster oocytes. Gamete Res 21:335-343.
> Johnson LA, Welch GR, Keyvanfar K, Dorfmann A, Fugger EF, Schulman JD
> (1993): Gender preselection in humans? Flow cytometric separation of X
> and Y spermatozoa for the prevention of X-linked diseases. Human Reprod
> 8:1733-1739.
> Morrell JM (1991): Applications of flow cytometry to artificial
> insemination: a review. Vet Rec 129:375-8.
> Pasteur X, Maubon I, Sabido O, Cottier M, Laurent JL (1992): Comparison
> of the chromatin stainability of human spermatozoa separated by
> discontinuous Percoll gradient centrifugation. A flow cytometric
> contribution. Anal Quant Cytol Histol 14:96-104.
> Pasteur X, Sabido O, Maubon I, Perrin CM, Laurent JL (1991):
> Quantitative assessment of chromatin stability alteration in human
> spermatozoa induced by freezing and thawing. A flow cytometric study.
> Anal Quant Cytol Histol 13:383-90.
> Peter AT, Jones PP, Robinson JP (1993): Fractionation of bovine
> spermatozoa for sex selection - a rapid immunomagnetic technique to
> remove spermatozoa that contain the H Y antigen. Theriogenology
> 40:1177-1185.
> Seligman J, Shalgi R, Oschry Y, Kosower NS (1991): Sperm analysis by
> flow cytometry using the fluorescent thiol labeling agent
> monobromobimane. Mol Reprod Devel 29:276-281.
> Spano M, Evenson DP (1993): Flow cytometric analysis for reproductive
> biology. Biology of the Cell 78:53-62.
> Tao J, Du JY, Critser ES, Critser JK (1993): Assessment of the
> acrosomal status and viability of human spermatozoa simultaneously
> using flow cytometry. Human Reprod 8:1879-1885.
> 
> 
> Jim Cummins                   
> School of Veterinary Studies
> Murdoch University
> Western Australia 6150  Tel +61-9-360 2668 Fax +61-9-310 4144
> "An inordinate fondness for Beetles"

####Just to set the record straight, the blunder about flow cytometery and
sperm was mine, and not Patrick Lamastus (sorry Patrick). -Will, U. of
Penn./Neurology



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