Flow sorting of X and Y chromosome‐bearing spermatozoa into two populations
作者:
L. A. Johnson,
J. P. Flook,
M. V. Look,
D. Pinkel,
期刊:
Gamete Research
(WILEY Available online 1987)
卷期:
Volume 16,
issue 1
页码: 1-9
ISSN:0148-7280
年代: 1987
DOI:10.1002/mrd.1120160102
出版商: Wiley Subscription Services, Inc., A Wiley Company
关键词: flow cytometry;DNA;sperm separation;fluorescent stain
数据来源: WILEY
摘要:
AbstractThe only established difference on which to base the separation of X and Y chromosome‐bearing spermatozoa is chromosomal constitution. This difference is quantifiable both from chromosome morphology (karyotype) and from DNA content. Flow cytometric techniques were used to measure relative DNA content of the X and Y populations and to flow‐sort spermatozoa fromChinchilla laniger.Epididymal spermatozoa were recovered in PBS, fixed in 80% ethanol, treated with papain and dithioerythritol, and stained for DNA with Hoechst 33342. Sperm nuclei were analyzed and sorted on an EPICS V flow cytometer/cell sorter, modified specifically for spermatozoa. Two clearly resolved peaks (coefficient of variation<1.5%) with approximately 7.5% difference in DNA content between X and Y chromosome‐bearing spermatozoa were evident. Sperm nuclei were sorted from a portion of the X and Y peaks at a rate of 55 nuclei/sec for each population. Purities of individual X and Y populations averaged 95% as determined by reanalysis of the sorted populations. Successful sorting ofChinchillaX and Y chromosome‐bearing spermatozoa into separate populations may aid in the identification of a biochemical marker that could be used to discriminate between the two sperm populations and lead to a practical procedure for sexing sper
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