I have a student (actually someone else's student)
 needing to examine sperm morphology: normal versus
abnormal, head details more important than tail.
We have tried
1. rhodamine dyes - give nice mitochondrial detail, but
tail and head do not stain enough to image clearly.
2. fluorescein diacetate - gave very weak signals, too low for
accurate morphology - makes me wonder what esterases
there are in sperm.
 
I haven't tried calcein yet - our next step.... but I can't afford to go
step by step through a battery of fluorophores, and the student is
getting a little desperate, so if anyone can suggest any probes that
may allow us to image and do 3D of living sperm,  please let me know.
Any comments and/or references would be greatly appreciated. BTW, we
immobilize the sperm in low temp gelling agarose to prevent movement
artfacts during scanning - no obvious morphological changes seem by
DIC.
 
Thanks
 
Ian
 
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Dr Ian Harper
Experimental Biology Programme
Medical Research Council
PO Box 19070                         Tel: 027-21-938 0347
Tygerberg 7505                       Fax: 027-21-938 0456
South Africa            Internet: [log in to unmask]
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