RE: sperm staining

From:Kathy Paton <>

Dear Deborah
I have used the following method with excellent results.
Bergs Stain for spermatozoa
Putts carbol-fuchsin
   New fuchsin            1gm
   Phenol                 5gm
   Distilled water       84ml
Absolute alcohol         10ml

Saturated Lithium Carbonate
   Lithium Carbonate      1gm
   Distilled water      100ml

Glacial acetic alcohol
   Glacial acetic acid    5ml
   Absolute alcohol      95ml

Methylene Blue
   Methylene blue        0.5gm
   Absolute ethanol      100ml


1   Sections to water
2   Stain for 3min in carbol fuchsin (filtered)   3 min.
3   Place in lithium carbonate for  3min
4   Decolourize in glacial acetic alcohol for 5min (control microscopically)
5   Wash in two 1-minute changes of abs alc
6    Counterstain in methylene blue for 30 to 60 seconds.
7   Rinse rapidly in a couple of changes of abs alc
8   Clear in several changes of Xylol...mount.

Spermatozoa   brilliant red
Erythrocytes  pale pink

Postive control is required.......this in itself can prove to be fun even in
a busy lab.
I will comment no more

Kathy Paton
Surgical Pathology Unit
North Shore Hospital
New Zealand
-----Original Message-----
From: Deborah Faichney []
Sent: Tuesday, November 07, 2000 4:54 AM
To: ''
Subject: sperm staining

Hello all,

A biology colleague would like to be able to stain sperm in the seminal
vesicle of whole soil mites. 
(I always seem to attract strange requests from colleagues, fish related
questions are bad enough, never mind soil mites!)

They have cleared them in lactic acid, and would love a method to stain the

Any suggestions??


Debbie Faichney
Snr. Histopathology Technician

Institute of Aquaculture
University of Stirling

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