PROCEDURE OF GIEMSA STAINING. The Procedure of Giemsa staining varies as per the purpose of staining that means whether the staining is done for the examination of Blood cells or to find the Parasites in the blood smear and accordingly the Blood smears are prepared as Thin Blood films or Thick blood films.
Giemsa stain is a buffered thiazine-eosinate solution designed to provide coloration of blood cells similar to the original product described by Giemsa. It may be used separately or in combination with a May Grünwald Stain, also available from Sigma-Aldrich. _____ _____REAGENT GIEMSA STAIN Catalog No. GS
GIEMSA STAINING OF MALARIA BLOOD FILMS MALARIA MICROSCOPY STANDARD OPERATING PROCEDURE – MM-SOP-07A 1. PURPOSE AND SCOPE To describe the procedure for properly staining malaria blood films with Giemsa stain. This procedure is to be modified only with the approval of the national coordinator for quality assurance of
Giemsa stain is also used to visualize chromosomes.This is particularly relevant for detection of Cytomegalovirus infection, where the classical finding would be an "owl-eye" viral inclusion.. Giemsa stains the fungus Histoplasma, Chlamydia bacteria, and can be used to identify mast cells.. Generation. Giemsa's solution is a mixture of methylene blue, eosin, and Azure B.
Staining: Place 1.0ml of the Wright-Giemsa Stain upon the smear, in sufficient quantity to cover the entire surface, for 3-4 minutes. Add 2.0ml distilled water or Phosphate Buffer, pH 6.5 and let stand twice as long as in step 1.
do not let the stain solution dry over the smear. Giemsa staining procedure . Fix the slide in methanol. Then pour or flood the slide with Giemsa stain and allow to react for 20 minutes; Wash off the smear with clean water; Allow to air dry and examine with 100x microscope magnification using immersion oil.
Nowadays, commercially prepared Giemsa staining solutions are used in most of the laboratories. However, some laboratories still prepare it in their own setup as per their Standard Operating Procedure (SOP). Check out the Preparation of Giemsa Stain or Giemsa Stock solution in Laboratory....
4. Stain the entire slide with diluted Giemsa stain (1:50, vol/vol) for 50 min. For a 1:50 dilution, add 2 ml of stock Giemsa to 40 ml of buffered water in a Coplin jar. Place the slide in the stain, thick film down to prevent the debris caused by dehemoglobinization from falling onto the thin film. 5.
One alternate is 10 minutes in 10% Giemsa; the shorter stains yield faster results, but use more stain and might be of less predictable quality. Staining Procedure: Quality Control. To ensure that proper staining results have been achieved, a positive smear (malaria) should be included with each new batch of working Giemsa stain.
Observe the results of the staining procedure under oil immersion (100x) using a Bright field microscope. If you are struggling to remember the staining reagents used in this procedure and their order you can remember this sentence “Come In And Stain” i.e. the order is Crystal violet, Iodine, Alcohol/Acetone and the final one is Safranin.