toxoplasma
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From: | Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
We found (worked with a Toxo guru!) that thin GMA sections worked
very well, but did a modified PASH,.
Routine paraffin sections were handled as described.
NBF fixation (Trumps 4% formalin/1% gluteraldehyde also excellent)
section at 5 um
I did not put all rinse steps in
hydrate,
0.5% periodic acid (freshly made each time ) 20 min
Schiffs reagent 20 miutes, followed by a 10 min running tap
water rinse to turn red.
Gill III hematoxylin 7 min( overstained with this)
clarify with 4% acetic acid 10 dips
Blue 2 min
rinse, dehydrate, mount.
Cysts are purple pink, nuclei blue.
You could try a Weigerts Iron hematoxylin instead, but the PAS was
important. Hardest thing is to get a good positive control,
and recognize the cysts in tissues, but with practice, this was
a standard stain for us. Also, overstaining with hematoxylin
seemed to help for H&E.
Good luck
Gayle Callis
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