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UND 363 Gridley

purpose for Gridley Fungus Stain demo fungi
principle chromic acid oxidizes adjacent glycol groups to aldehydes. Aldehydes then react to schiff reagent.
how is this principle different than the PAS stain chromic acid is a stronger oxidizer than periodic acid and will destroy/turn some aldehydes to acid therefore leaving less reactive groups to react with schiff. (less intense reaction than PAS but also less background staining.
In addition to the chromic acid destroying many aldehyde groups what will the aldehyde fuchsin do in its place. the aldehyde fuchsin will act as an aldehyde and occupy uninvolved linkages of the schiff reaget thereby reinforcing the depth of the stain.
fix NBF
micron 4-5
QC section with fungus
reagents in order 4% chromic acid - oxidizer, Schiff - reacts with aldehydes, aldehyde fuchsin - acts as aldehyde by occupying uninvolved linkages of schiff reagent, metanil yellow - counterstain
results mycelia/elastic fibers/mucin - deep purple, conidia - deep rose to purple, background - yellow
how will old non-viable fungi be stained with this technique? they are not well stained if they are non-viable
what type of acid can be used after the schiff reagent sulfurous acid - will remove excess schiff reagent
of pararosaniline (CI42500) and basic fuchsin (rosaniline CI 42510) which is use to prepar the aldehyde fuchsin pararosaniline
explain in detail what the chromic acid will do the glycols (in trace amts. Vs. those with heavy polysaccharide %) oxidizes 1,2 glycols to aldehydes and then oxidizes some further to acid (trace amts.) Tissue with heavy polysaccharides (mucin, glycoten, fungi) take longer and will still show + PAS RXN even after basement membrane and conn. tissue are non-reactive.
Created by: mustangvxd



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