- Catalog Order # K7024 - 4 oz. kit -$61.00
- K7028 - 8 oz. kit - $101.00
APPLICATION
Mammalian Tissue
DESIGNED TO SHOW
Gram Positive and Negative bacteria
SPECIMEN REQUIREMENTS
Fixation: Tissue: Any well fixed tissue , however, it is suggested to use formalin fixed tissue.
Fixation; Smears: Air dried and alcohol fixed.
Sections; Cut Paraffin sections at 6 microns.
COMPONENTS AND REAGENTS
1. Gentian Violet Solution - 4oz / 120ml
2. Gram?s Iodine Solution - 4oz / 120ml
3. Safranin Stain Solution - 4oz / 120ml
4. Picric Acid/Acetone - 4oz / 120ml
5. Gram?s Decolorizer Solution - 4oz / 120ml
6. Control Slides1 Stained and 3 unstained
7. Pap Jars
STAINING PROCEDURE
1. Deparaffinize sections and hydrate to deionized water.
2. Stain sections in Gentian Violet Stain for 30 seconds.
3. Wash sections in running tap water until excess stain is removed.
4. Treat sections in Gram?s Iodine for 1 minute.
5. Rinse sections thoroughly under tap water.
6. Treat sections in Gram?s Decolorizer to remove stain until background is translucent. 7. Rinse sections quickly under tap water.
8. Stain sections in Safranin Stain for 2 minutes.
9. Wash sections quickly in tap water.
10. Dip slides in Picric Acid/Acetone 6 times.
11. Immediately blot slides and allow to air dry.
12. Clear slides in two changes of Xylene or Xylene Substitute.
(Increase time in Xylene Substitutes)
13. Mount using Clearslip.
RESULTS
Gram Positive Cocci-Black/Blue
Negative Bacilli-Red
NucleiRed
BackgroundYellow
NOTES
1. Do not extend treatment in Picric Acid/Acetone if Safranin stain has been removed totally from section. Fanning the slide will assist in the evaporation of acetone in the Picric Acid/Acetone. The longer the Acetone is on the slide, the more Safranin that will be differentiated from section.
REFERENCES
1. AFIP, Manual of Histologic Staining Methods, American Registry of Pathology, 1992
2. Luna LG: Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts, Gaitherburg, MD, American Histolabs, Inc., 1992
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