RE: [Histonet] GRAM Stain Timing Question

From:"Owen, Michael P"

Dear Sara,

In my laboratory personnel when using Coplin jars Gram staining is
usually performed as below. I have good results most of the time
following this procedure.

Crystal Violet: 60 seconds
Tap Water Rinse
Iodine: 60 seconds
Tap Water Rinse
50% w/v Acetone-95% Ethanol: 40 seconds
Tap Water Rinse
Safranin: 60 seconds
Tap Water Rinse and Air Dry

My laboratory usually has Remel or Difco/BD Gram stain kits. The
decolorizer runs out quickly therefore I often make my own that is
equivalent to ones found in these kits.



Michael P. Owen, Regulatory Microbiologist
U.S. FDA Pacific Regional Lab Northwest
22201 23rd Drive SE  Bothell, WA 98021-4421
Phone: 425-483-4865     E-Mail:

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