Chromosome Immuno-Histo chemistry
Christopher R. Cederroth
Thursday, 14 August 2008 14:27 UTC
Hello!
I am trying to identify pachytene-stage spermatocytes from mice testis using the nuclear spread technique (Hypotonic shock with sodium citrate 1%, 2×10’ at 37°C; fixation with 3/4 methanol, 1/4 acetic ac., 10’ at RT; rinse with acetic ac. 70%, spread on slide, dry and stain with Gymsa ).
I have a few questions :
What does a early, mid, late pachytene do look like?
Why do people use silver nitrate in addition to Gymsa when assessing for pachytene integrity? I think it stains for NOR, but I don’t understand what other info it brings.
Can I perform immuno stainings on samples fixed as described above?
Thanks for your help!
Chris
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