RE: sperm staining
From: | Kathy Paton <Patonk@WHL.co.nz> |
Dear Deborah
I have used the following method with excellent results.
Bergs Stain for spermatozoa
SOLUTIONS
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
METHOD
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.
RESULTS
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
Auckland
New Zealand
-----Original Message-----
From: Deborah Faichney [mailto:d.a.faichney@stir.ac.uk]
Sent: Tuesday, November 07, 2000 4:54 AM
To: 'histonet@pathology.swmed.edu'
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
sperm.
Any suggestions??
Thanks
Debbie Faichney
Snr. Histopathology Technician
Institute of Aquaculture
University of Stirling
Stirling
Scotland
UK
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