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UND 363 Kinyoun AFB

UND 363 Kinyoun Acid Fast

QuestionAnswer
purpose for Kinyoun Acid Fast (A/F)stain 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
Why is heat necessary (Vs. room temp) for this technique lipoid capsule is waxy at room temp, the heat allows the dye to penetrate into the bacteria
fix for Kinyoun Acid Fast stain NBF preferred, others can be used EXCEPT CARNOYS WHICH CANNOT BE USED BECAUSE IT WILL CAUSE A/F BACTERIA TO BECOME NON A/F
what does Carnoys fixative do to A/F bacteria It will make them NON A/F
microns 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.
why can tap or distilled water NOT BE USED before the carbol-fuchsin step there may be bacteria in the piping
Reagents in order for Kinyoun Acid Fast stain Millipore water, kinyoun carbol fuchsin (basic fuchsin/phenol crystal/alcol), HCL/alcohol - diff'r, methylene blue - counterstain
in the Kinyoun carbol fuchsin what does the carbol fuchsin do? what about the Phenol/alcohol? C.F. stains the lipid capsule, the phenol and alshol enhances staining and aids in dissolving of the stain.
results for Kinyoun Acid Fast stain A/F bacteria - bright red, background - blue
what must be done after the adic alcohol step (hcl/alcohol) must be rinsed well before counterstaining or the tissue will not stain
why can the sections not be allowed to dry out after carbol fuchsin a compound that resists decolorization will form
what helps the penetration of phenol (AKA carbolic acid) in this stain heat and the action of wetting agents (which reduce surface tension and enhance spread of phenol)
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 (step 4) to remove methylene blue, wash with water and repeat counterstain
what can kinyoun A/F not be used for and why mycrobacterium leprae (Must use the FITE stain instead). This bacteria is sensitive to alcohols.
Created by: mustangvxd