McDonald Gram Procedure

From:Cindy/Rick DuBois

Below is the procedure for the McDonalds Gram Stain.  All credit for this technique goes to American Mastertech.  It is a staining kit they prepare.  They can be reached via internet:  www.mastertechs.com, or phone (800)860-4073.  The item number for the kit is KTMGS.

1.  Flood slides for 1  minute with Gentian violet.
2.  Rinse in water.
3.  Flood slides with Gram's Iodine, let stand 1 minute.
4.  Rinse.
5.  Decolorize in Gram's decolorizer until color no longer                bleeds off of slide.
6.  Stain with Carbol Fuchsin for 90 seconds (we find that 60 seconds works just as well).
7.  Rinse quickly.
8.  Stain with Tartrazine Solution for 15 seconds, (0ur docs like lots of yellow for background so we stain 30 seconds).
9.  Dehydrate quickly, clear thru xylene or xylene substitute and coverslip.

RESULTS:
 GRAM POSITIVE:  BLUE
 GRAM NEGATIVE:  RED
 OTHER TISSUE:   YELLOW
  NUCLEI:        RED.

Have fun with this,
Cindy DuBois, HT
Delta Pathology Associates
Stockton, CA  95207
(209)477-4432



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