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UND 363 HotchkissPAS

UND 363 Hotchkiss PAS

QuestionAnswer
principle for Hotchkiss PAS demo of fungi
principle for Hotchkiss PAS polysaccharides in cell walls are oxidized with periodic acid to aldehydes. The aldehydes react with Schiff to create a rose colored fungi
fix NBF, bouin, zenker
micron 4-5
QC section with fungi
reagents in order periodic acid - oxidizer, schiff reagent - react with aldehys for pink color, sulforous rinse - remove excess schiff and prevent false colorization, fast green 1:5000 - counterstain
results fungi - rose, background - green
if there is a lack of contrast and weak background what is the issue and what can be done? the oxidizing soln' (periodic acid) is weak therefore it will not oxidize native carbs past the aldehyde stage which in turn allows these elements and carb rich fungus to react with schiff, FIX - use strong oxidizer (ie chromic acid)
what is another reason there is a lack of contrast and weak background what is the issue and what can be done? the schiff reagent is old and depleated, FIX - use fresh
if there is weak staining/fungus not staining what can the problems be and the FIX Exhausted oxidizer - FIX use fresh, Overused schiff reagent will produce poor results - FIX use fresh
Created by: mustangvxd