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UND 363 Brown Hopps

UND 363 Brown Hopps Gram

purpose For Brown Hopps Gram Stain Mod Demo gram negative and gram positive bacteria
Principle For Brown Hopps Gram Stain Mod (4 parts) Crystal violet stains all cells and tissue blue, Iodine is added as a mordant (forming dye lake), alcohol or acetone decolorizes gram neg. bacteria, counterstain applied for gram neg.
What is the reason that the gram negative cells lose color due to alcohol or acetone Gram positive have thicker cell wall of peptidoglycan whereas gram negative cell walls are thinner ( 15-25 Vs. 8-12). The alcohol disrupts the the outer lipoprotein layer and it cannot retain the violet-iodine complex
Why will gram positive retain the large crystal violet-iodine complex The INTACT cell walls are thicker and can resist decolorization from alcohol or acetone (unless it has been damaged)
How is the Brown Brenn Gram stain different in reference to alcohool and acetone it uses equal parts acetone and alcohol to decolorize
how is the gram twort gram stain different in regards to counterstaining it uses a combonation of neutral red and fast green as a counterstain
fix For Brown Hopps Gram Stain Mod NBF, but 2.5 buffered gluteraldehyde or hellys ok
which fix is not ok For Brown Hopps Gram Stain Mod Bouins
microns 4-5
QC section with both gram positive and gram negative
reagents in order 6 crystal violet - stain gram positive, grams iodine - mordant (iodine and potassium iodide), acetone - decolorize, basic fuchsin - cntrstain for gram neg., gallego soln - diff'r, picric acid/acetone - decolor and background stain
what makes up the gallego differentiation soln formalin and acetic acid
results For Brown Hopps Gram Stain Mod gram pos. - blue, gram neg. - red, background - yellow, nuclei - light red.
what are the results if the gram twort mod of counterstain is used gram pos. - blue/black, gram neg. - red/pink, nuclei - red, collagen/rbc/cytoplasm - shades of green
what can cause the gram positive to not stain (per reagent use) and fix stained slides were in the alcohol tool long (step before clearing and coverslipping) - quickly dehydrate through alcohol to xylene IE overdecolorized
what can cause the gram positive to not stain (per patient) and fix bacteria are dead or treated with antibiotics fix - increase time in crystal violet and iodine OR reduce time in decolorizer OR use a less potent differentiation.
what can cause gram positive bacteria not to stain (per solvents) (part of step 3), over decolorized with organic solvent. THIS IS THE MOST COMMON REASON THE STAIN DIDN'T WORK. Fix - reduce time in solvent and wash w/ H20 OR use slower diffr' (alchohol/acetone mix or alcohol instead of acetone, or h20 diluted acetone)
give 2 reasons that the background would stain blue black and fis underdecolorzed with organic solvent, FIX - increase time in decolorizer or use quicker solvent (ie acetone). Crystal violet dried and created and insoluble compound. FIX - don't allow to dry.
Give 2 reasons that gram Negative bacteria won't stain dehydration (step 4) after red dye was too long - FIX - reduce time in dehydrants. Crystal violet was not fully removed (step 3) FIX - increase time in decolorizer.
Give 3 reasons that gram neg. bacteria wont stain (exclude dehydration time error, and crystal violet retention errors) Not enough time in dehydrant/gallego soln. FIX - watch gallego diff'r (gallego leaches red dye from background. Counterstain was NOT used. Error with red counterstain (increase time in red dye/prepare fresh
what can the Brown hopps gram stain also be used for cryptcococcus, aspergillosis, rhinosporidiosis, amebiasis, actinomycosis, blastomycosis,
Created by: mustangvxd



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