Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-30T22:21:23.951Z Has data issue: false hasContentIssue false
  • English
  • Français

Counting dead spermatozoa in frozen semen

Published online by Cambridge University Press:  02 September 2010

H. R. L. Buttle ,
J. L. Hancock  and
A. F. Purser
H. R. L. Buttle
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
J. L. Hancock
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
A. F. Purser
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
Get access

Summary

Three methods were used to make differential counts of living and dead bull spermatozoa in samples of frozen semen in an egg-yolk citrate medium containing glycerol.

With two methods (‘phase’ and ‘nigrosin’ methods) dead spermatozoa were identified by their altered structure. With a third method (nigrosineosin) dead spermatozoa were identified by their staining affinity. With the phase method spermatozoa were immobilised by treatment with approximately M/40 sodium fluoride and were examined by phase-contrast microscopy in unfixed wet preparations. With the other two methods the spermatozoa were examined in smears stained either with nigrosin alone (nigrosin method) or with nigrosin and eosin (nigrosin-eosin method).

Analysis of variance of the results of a factorial experiment involving fluoride-treated and untreated samples from 6 bulls with the three methods showed that differences between semen samples contributed 66% of the variation. A defect of the phase method was that the variance between counts was greater than the theoretically expected value.

All spermatozoa in nigrosin-eosin stained preparations were stained with eosin within a few days of the smears being made, so that living and dead spermatozoa could not be distinguished by their differing affinities for eosin. Repeat counts on nigrosin-stained preparations did not differ significantly from counts made several days previously. Sodium fluoride in the concentration used here (M/40) tended to reduce the percentage of dead spermatozoa.

Type
Research Article
Information
Animal Production , Volume 7 , Issue 1 , January 1965 , pp. 59 - 65
Copyright
Copyright © British Society of Animal Science 1965

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Beatty, R. A., 1957. Nigrosin-eosin staining of rabbit spermatozoa and the fertility of semen. Proc. roy. Soc. Edin. (B), 67: 1.Google Scholar
Beatty, R. A., & Napier, R. A. N., 1960. Genetics of gametes. I. A quantitative analysis of five characteristics of rabbit spermatozoa. Proc. roy. Soc. Edin. (B), 68: 1.Google Scholar
Blackshaw, A. W., 1958. The effects of glycerol on the supra-vital staining of spermatozoa. Aust. Vet. J., 34: 77.CrossRefGoogle Scholar
Campbell, R. C., Dott, H. M., & Glover, T. D., 1956. Nigrosin-eosin as a stain for differentiating live and dead spermatozoa. J. agric. Sci., 48: 1.CrossRefGoogle Scholar
Campbell, R. C., Hancock, J. L., & Rothschild, Lord, 1953. Counting live and dead bull spermatozoa. J. exp. Biol., 30: 44.CrossRefGoogle Scholar
Campbell, R. C., Hancock, J. L., & Shaw, I. G., 1960. Cytological characteristics and fertilising capacity of bull spermatozoa. J. agric. Sci., 55: 1.CrossRefGoogle Scholar
Fisher, R. H., & Yates, F., 1949. Statistical Tables for Biological, Agricultural and Medical Research. Oliver & Boyd, Edinburgh.Google Scholar
Hancock, J. L., 1952. The morphology of bull spermatozoa. J. exp. Biol., 29: 445.CrossRefGoogle Scholar
Hancock, J. L., 1957. The morphology of boar spermatozoa. J. roy. Micr. Soc, 76: 77.CrossRefGoogle ScholarPubMed
Mixner, J. P., & Saroff, J., 1954. Interference of glycerol with differential staining of bull spermatozoa as used with semen thawed from the frozen state. J. Dairy Sci., 37: 1094.CrossRefGoogle Scholar