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UND 363 Ziehl/Neelse

UND 363 Ziehl/Neelsen

QuestionAnswer
purpose for Ziehl-Neelsen (SAME AS KINYOUN EXCEPT FOR A FEW DIFFERENCES) demo of acid fast mycobacteria
principle lipoid capsule of A/F mycobacteria takes up carbol fuchsin and resists decolorization with dilute mineral ACID.
what is carbol fuchsin more soluble in (lipids of cell wall or acid-alcohol) and how does this affect NON A/F bacteria it is more soluble in the lipids of the cell wall. NON A/F bacteria lack the waxy capsule and therefore carbol-fuchsin is easily removed
what does the Phenol and alchol in this stain do? they aid in dissolving the basic fuchsin
fix any well fixed tissue except CARNOYS
what does Carnoys fixative do to A/F bacteria It will make them NON A/F
micron 4-5
Qc tissue with A/F organisms, must use Millipore filtered (.45 micron pore size) water, and a negative control from same day workload.
what are the DIFFERENCES from the Kinyoun technique stain times for carbol fuchsin (30 min. Vs. kinyoun 1hr.), and conc. % of reagents (Phenol/ABSOLUTE/fuchsin 2.5ml/5ml/.5g) Vs. (kinyoun 56ml/95% ALCOHOL 140ml/28g)
if overstained with methylene blue what will occur and what can be done it will mask any organisms present. Fix - take slide back to acid alcohol to remove methylene blue, wash with water and repeat counterstain
why can the sections not be allowed to dry out after carbol fuchsin a compound that resists decolorization will form
results A/F bacteria - bright red, background - lt. blue
why can tap or distilled water NOT BE USED before the carbol-fuchsin step there may be bacteria in the piping
what can Ziehl-Neelsen not be used for and why mycrobacterium leprae (Must use the FITE stain instead). This bacteria is sensitive to alcohols.
what must be done after the adic alcohol step (hcl/alcohol) must be rinsed well before counterstaining or the tissue will not stain
in the Ziehl-Neelsen what does the carbol fuchsin do? what about the Phenol/alcohol? C.F. stains the lipid capsule, the phenol and alcohol enhances staining and aids in dissolving of the stain.
Created by: mustangvxd
 

 



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