Giemsa

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining spleen.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  May-Grunwald Giemsa 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:

APPLICATION:

Newcomer Supply Giemsa Control Slides are for the positive histochemical and differential staining of hematopoietic tissue.

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. 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.
      1. 1. See Procedure Notes #1 and #2.
   3. Rinse in two changes of Alcohol, Methanol Anhydrous, ACS (Part 12236); 3 minutes each.
   4. Prepare fresh Working Jenner Stain Solution just prior to use; combine and mix well.
      1. 1. Distilled Water       20 ml
         2. Jenner Stock Stain                       20 ml
   5. Place slides in fresh Working Jenner Stain Solution for 6 minutes.
   6. Prepare fresh Working Giemsa Stain Solution just prior to use; combine and mix well.
      1. 1. Distilled Water       47 ml
         2. Giemsa Stock Stain, Wolbach         3 ml
   7. Place slides directly into fresh Working Giemsa Stain Solution for 45 minutes.
   8. Rinse quickly in distilled water.
   9. Differentiate each slide individually in Acetic Acid 1%, Aqueous (Part 10012); 6-10 dips.
      1. 1. Check microscopically for well differentiated nuclei.
   10. Rinse in distilled water.
   11. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

RESULTS:

PROCEDURE NOTES:

1. 1. Drain slides after each step to prevent solution carry over.
   2. Do not allow sections to dry out at any point during procedure.
   3. The color range of the stained cells may vary depending upon fixation and degree of differentiation.
   4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

REFERENCES:

1. 1. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 121-122.
   2. Shapiro, Stanley H. and Hilda Laufer. “Observations on Fixation and Staining of Bone Marrow Biopsies.” The Journal of Histotechnology3 (1988): 145-47.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 157.
   4. Modifications developed by Newcomer Supply Laboratory.