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UND 363 Mwave MSN

UND 363 Mwave Methenamine Silver Nitrate

QuestionAnswer
purpose for Mwave Methenamine Silver Nitrate rapid diagnosis of P jirovecii and pneumocystis corinii on cytospin preps or F/S's
Principle for Mwave Methenamine Silver Nitrate Fungal cell wall polysaccharides are oxidized to aldehydes by chromic acid. Cell walls/glycogen/mucins with large % polysacc.'s will remain reactive to methenimine-silver and reduced to visible.
fix cytospin preps fixes in 95% alcohol, FS fixed in 37-40% formaldehyde, standard paraffin sections with NBF
microns FS - 6 micron, paraffin - 4-5, **sections tend to wash off with this stain so charged slides should be used (poly l lysine or silanized slides)
QC use a control with the fungus you are looking for. chem. cleaned glassware and nonmetal forceps needed
reagents in order 10% chromic acid - oxidizer, soduim bisulfite 1% - remove chromic acid/clearing agent, methenimine silver - impregnation, gold chloride - toner, soduim thiosulfate - remove unreduced silver, lt green -counterstain
results fungi and P jirovecii cell wall - black, background - Green
how should the plastic coplin jars be used in the Mwave put the plastic jars in closed plastic bags, open microwaved bags under fume hood OR put mwave under fume hood
if slides are left too long in methenamine silver what will occur reticular fibers, RBCs and other tissue may be stained. The non specific staining may mimic fungi or obscure small fungi leading to potential false positives.
What is mirroring, what can cause it, FIX it? random deposits of silver onto slides. caused by over heating or dirty glassware. FIX - don't overheat, use ammoniacal silver for impregnation.
Created by: mustangvxd