Laboratory
(BHHRL)
Tel: +267 3902671;
Fax: +267 3901284 / Standard Operating Procedure
Quality Control of May-Grunald Giemsa Stain / Document: BHHRL/005PR02
Version: 1
Author: See Below / Document Number: / Pro64-B-12
Effective (or Post) Date: / 17-Jan-08
Document Origin: BHHRL / Company: / NA
SMILE Approved by: / Heidi Hanes
Review by / Heidi Hanes / Review date / 7 Feb 2012
SMILE Comments: This document is provided as an example only. It must be revised to accurately reflect your lab’s specific processes and/or specific protocol requirements. Users are directed to countercheck facts when considering their use in other applications. If you have any questions contact SMILE.
Title: / Quality Control of May-Grunald Giemsa Stain
Prepared By: / Name, Title / Signature, Date
Munyaradzi P. Mangwendeza, QA Manager
Approved By: / Name, Title / Signature, Date
Dr. Rosemary Musonda, Laboratory Director
Annual Review / Review Date / Revision Date / Signature, Date
Distributed To: / Location: / Copy Number: / Location: / Copy Number
Master file / 1
Director –
Dr. Musonda / 2
Director –
Dr. Mine / 3
Lab Managers – Mr. S. Moyo
Mrs. K. Makhaola / 4
5
Lab Coordinator / 6
Copy Number: ______
1. General Principle
1.1. All reagents used at BHHRL should go under quality control as prescribed to ensure that they continue to perform as expected all the time.
2. Purpose
2.1. To give directions on how to carry-out quality control activities on the May-Grunald Giemsa Stain used for the manual white cell differential counts.
3. Scope
3.1. The SOP covers quality control activities associated with the May-Grunald Giemsa Stain.
4. Responsibilities
4.1. Haematology lab staff members involved in differential white cell counts are to follow the directions of the SOP to ensure that their activities in manual differential white cell counts produce reliable and correct results.
5. Procedures
5.1. Giemsa stain is used to differentiate nuclear and/or cytoplasmic morphology of platelets, RBCs, WBCs, and parasites.
5.2. Liquid stock is available commercially. The stain must be diluted for use with water buffered to pH 6.8 or 7.0 to 7.2, depending on the specific technique used. Either should be tested for proper staining reaction before use.
5.3. The stock is stable for years, but it must be protected from moisture because the staining reaction is oxidative. Therefore, the oxygen in water will initiate the reaction and ruin the stock stain. The aqueous working dilution of stain is good only for 1 week.
5.4. The stock buffer solutions and buffered water should be clear, with no visible contamination.
5.5. Check the Giemsa stain reagents, including the pH of the buffered water, before each use.
5.6. Filter the stains before use each time.
5.7. Use the QC form to keep records of the Lot numbers of the reagents as well as the expiry dates.
5.8. Prepare and stain films from “normal” blood, and microscopically evaluate the staining reactions of the RBCs, platelets, and WBCs; this assessment can also be accomplished by the examination of patient slide. If the staining reactions are acceptable, then the QC is considered acceptable.
5.9. Macroscopically, blood films appear purplish. If blue, the buffered water was too alkaline; if pink to red, the buffered water was too acid.
5.10. Microscopically, RBCs appear pinkish gray, platelets appear deep pink, and WBCs have purple-blue nuclei and lighter cytoplasm. Eosinophilic granules are bright purple-red, and neutrophilic granules are purple. Basophilic stippling within uninfected RBCs is blue.
5.11. Slight variation may appear in the colors described above depending on the batch of stain used and the character of the blood itself, but if the various morphological structures are distinct, the stain is satisfactory.
5.12. Discard used reagents appropriately at the end of each week.
5.13. Clean jars with mild soap and water and always rinse with de-ionized water.
5.14. The microscope should be recalibrated once each year. This recommendation should be considered with heavy use or if the microscope has been bumped or moved multiple times.
5.15. Record all QC results on the QC form BHHRL/005FR01. The completed forms should be reviewed at the end of each month by the supervisor.
6. Records
6.1. Stain QC records.
7. Co-Applicable Document
7.1. BHHRL/005PR01 Manual Differential Cell Count.
8. References
8.1. Reagent Package inserts.
8.2. Garcia, L.S. 1999. Practical Guide to Diagnostic Parasitology, ASM Press, Washington, D.C.