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UND 363 Alk Congo Re

UND 363 Alkaline Congo Red

QuestionAnswer
Purpose of Alkaline Congo Red demonstrate amyloid
principle of Alkaline Congo Red congo red is linear molecule which allows for hydrogen bonding of the AZO and Amine groups of Amyloid
What is an underlying technique of Alkaline Congo Red per pretreatment w/ alkaloid alkaloid pretreatment to release native internal hydrogen bonds between protein chains and therefore more sites for dye binding
fixatives of Alkaline Congo Red Alcohol, or carnoy PREFERRED, but 10% NBF , bouins, zenkers are OK. ***Prolonged storaged in 10%NBF will decrease staining***
thickness and why 8-10microns, if not at this thickness the applegreen birefringence will not show
explain the specificity of the control for this stain the control need to be known (one that has been shown to contain amyloid)
explain some caveats with the control slides keep minimal on hand (increased age will decrease stain intensity), ALSO large deposits give less intense stain reaction than new small deposits
microscope of Alkaline Congo Red light and polarized scope (to ID amyloid)
what are the 3 major reagents of Alkaline Congo Red harris hematox - stains nuclei, alkaline salt soln' - releases hydrogen bonds, congo red- has AZO and amine groups that form hydrogen bonds with hydroxyl radicals of amyloid
typical results of Alkaline Congo Red amyloid - deep pink to red (light microscope)/apple green (w/ polarized microscope) elastic - pale pink (light micro) nuclei- blue
explain why congo red is better than other methods to remove false positives other methods can create false positive (and false birefringment) due to excess dye retained in tissue ***congo red will not give false pos. when done CORRECTLY
what components help give a clean background with congo red sodium chloride and high alcohol found in dye solvent depress dye ionization and acid based staining resulting in stained section with clean background
name to components of the stain that are extremely important saturation of solution is extremely important, and the thickness (ie too thin = faint red, too thick and you get yellow birefringence)
Created by: mustangvxd
 

 



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