Supporting Laboratory Techniques > Basic Measurement Techniques

User Ratings

Your rating: None
Your rating: None
Your rating: None
 Add your comments

Epididymal Sperm Count

Yefan Wang1

1TherImmune Research Corporation, Gaithersburg, Maryland

Publication Name: 
Current Protocols in Toxicology
Unit Number: 
UNIT 16.6
DOI: 
10.1002/0471140856.tx1606s14
Print Publication Date: 
November, 2002
Online Posting Date: 
February, 2003
GO TO THE FULL TEXT:
PDF or HTML at Wiley Interscience
Are you the author of this protocol? Login or register and return to this page.

Abstract

Making Epididymal Sperm Counts (Yefan Wang, TherImmune Research Corporation, Gaithersberg, Maryland). Epididymal sperm counts are a widely used, simple and sensitive method of assessing the effects of male reproductive toxicants on the epididymal and/or testicular site of action. After careful dissection of the tissues and further processing, the sperm suspensions are counted using a hemacytometer and analyzed for effect.

     
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Interscience

Table of Contents

  • Basic Protocol
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Interscience

Materials

 Basic Protocol
 Materials
  • PBS (appendix 2A)
  • Necropsied male rats or mice, treated and control, euthanized
  • 15 × 60–mm (rats) or 15 × 30–mm (mice) and 1.5 × 2.0–cm petri dishes
  • Slide warmer
  • 13 × 100–mm test tubes and appropriate rack
  • Microscope: 10× and 40× objectives and 10× or 15× eyepieces
  • Necropsy board
  • 0.1-mm-deep improved Neubauer hemacytometer and coverslips
  • No. 11 scalpel blades
  • 60°C water bath
  • Gauze
  • Hand tally counter
  • Data collection system (i.e., hand-generated data form or computer-generated data collection system)
  • Additional reagents and equipment for using a hemacytometer (appendix 3B)

NOTE: All protocols using live animals must first be reviewed and approved by an Institutional Animal Care and Use Committee (IACUC) and must follow officially approved procedures for the care and use of laboratory animals.

Looking for materials?  Suppliers Guide
     
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Interscience

Figures

  • Figure 16.6.1
    A rat epididymis and its three anatomic parts: caput, corpus, and cauda.

    View Image
  • Figure 16.6.2
    Actual microscopic photograph of the secondary square (area within the triple line border) of a hemacytometer (see Fig. A.3B.1 in appendix 3B). The black arrows indicate the nine sperm that will be counted; five white arrowsidentify the sperm that will not be counted.

    View Image

Literature Cited

Literature Cited
    Hamilton Thorne Research 1989. HTM-IVOS Toxicology Application Version 10.8 Operation Manual. Hamilton Thorne Biosciences, Beverly, Mass.
    Kempinas, W.G. and Lamano-Carvalho, T.L. 1988. A method for estimating the concentration of spermatozoa in the rat cauda epididymis. Lab. Anim. 22:154-156.
    Klassen, C.D. 1996. Fifth Casarett & Doull's Toxicology: The Basic Science of Poisons, Fifth Edition. McGraw-Hill, New York.
    Saksena, S.K., Lau, I-F., and Chang, M-C. 1979. Age dependent changes in the sperm population and fertility in the male rat. Exp. Aging Res. 5:373-381.
    Ratnasooriya, W.D. and Wadsworth, R.M. 1987. Effect of mating on sperm distribution in the reproductive tract of the male rat. Gamete Res. 17:261-266.
     
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Interscience
Looking for Answers?
Do you have tips, tricks, or improvements to share?

Join the Conversation

Post new comment

The content of this field is kept private and will not be shown publicly.
CAPTCHA
This question is for testing whether you are a human visitor and to prevent automated spam submissions.