Gram Positive & Gram Negative Bacteria, Animal
Gram (+) and Gram (-) In animal tissue with specific infection or disease.
Tissue: Positive staining animal tissue and negative staining human lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain: Brown-Hopps quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.
Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.
CONTROL SLIDE VALIDATION:
|With Gram, Brown-Hopps Stain Kit:||Part 9124A||Individual Stain Solution|
|Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps||250 ml||Part 1041|
|Solution B: Iodine, Gram, Aqueous||250 ml||Part 1140|
|Solution C: Basic Fuchsin Stain 0.25%, Aqueous||250 ml||Part 1011|
|Solution D: Gallego Solution||250 ml||Part 1098|
|Solution E: Picric Acid-Acetone 0.05%||250 ml||Part 13351|
|Solution F: Acetone-Xylene 1:1||250 ml||Part 10015|
|Acetone, ACS||Part 10014|
Newcomer Supply Gram Positive & Gram Negative Bacteria, Animal Control Slides are for the positive histochemical staining of gram positive and gram negative bacteria in a naturally occurring infection.
NEWCOMER SUPPLY VALIDATION PROCEDURE:
- Heat dry sections in oven according to your laboratory protocol.
- Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
- See Procedure Notes #1 and #2.
- Stain in Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps for 2 minutes.
- Rinse well in distilled water, ensuring excess stain is removed.
- Mordant in Solution B: Iodine, Gram, Aqueous for 2 minutes.
- Sections should turn black.
- Rinse well in running tap water, ensuring excess iodine is removed.
- Blot excess water from slide; decolorize one slide at a time in Acetone, ACS (Part 10014) until blue color stops running; 1-2 dips.
- Sections should be very light gray in color.
- Quickly rinse in running tap water to remove excess Acetone.
- Stain in Solution C: Basic Fuchsin Stain 0.25%, Aqueous; 5 minutes.
- Rinse well in running tap water.
- Differentiate in Solution D: Gallego Solution for 5 minutes.
- Rinse in running tap water. Blot water off slide(s), but not to dryness.
- Proceed with Steps #13 to #16 one slide at a time.
- Dip quickly in Acetone, ACS (Part 10014); 1-2 dips.
- Dip directly in Solution E: Picric Acid-Acetone 0.05%; 3-10 dips.
- Dip quickly in Solution F: Acetone-Xylene 1:1; 5 dips.
- Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
|Gram positive bacteria||Blue/violet|
|Gram negative bacteria||Red|
|Nonreactive lung||Negative for gram positive/negative bacteria|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Brown, Robert C., and Howard C. Hopps. “Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.” American Journal of Clinical Pathology 60.2 (1973): 234-240.
- Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 222-224.
- Modifications developed by Newcomer Supply Laboratory.