Popular Answers (1)

The two methods for staining with Giemsa stain are the rapid (10% stain working solution) and the slow (3% stain working solution) methods. The rapid (10% stain working solution) method This is the commonest method for staining 1–15 slides at a time.

Giemsa stain preparation

1. Dissolve 3.8g of Giemsa powder within 250ml of methanol.
2. Heat the solution at 60oC.
3. Add 250ml of glycerin.
4. Filter the solution.
5. leave the solution to stand for about 1-2 months before use. Store the solution in a cool, dark place.

Make up a 10% Giemsa solution with distilled/deionized water buffered to pH 7.2. If only one slide is to be stained, you will require about 3 ml of prepared stain. Allow 3 drops of stock Giemsa solution (from the Pasteur pipette) to each millilitre of buffered water to give a 10% solution.

PRINCIPLE: The “neutral” dyes combining the basic dye methylene blue and the acid dye eosin, give a wide color range when staining. The pH of the staining solution is critical and ideally should be adjusted for different fixatives.

1. Dissolve 3.8g of Giemsa powder into 250ml of methanol.
2. Heat the solution from step 1 to ~60oC.
3. Slowly add in 250ml of glycerin to the solution from step 2.
4. Filter the solution from step 3.
5. The solution needs to stand a period of time prior to use.

May-Grunwald-Giemsa staining method is used for morphological inspection and differential counting of blood cells. May-Grünwald staining combines the effect of acidic eosin and alkaline methylene blue. Giemsa staining makes effect of azure. This staining stains all cellular components.

May-Grünwald is alcohol based and contains May-Grünwald’s eosin methylene blue and methanol (>85%). Giemsa is alcohol based and contains Giemsa’s azure eosin methylene blue, methanol (>50%) and glycerol.

Method

1. Dissolve 0.3 g of May Grunwald dye in 100 mL absolute methanol in a 250 mL conical flask.
2. Warm the mixture to 50°C in a water bath for a few hours and allow it to cool to room temperature.
3. Stir the mixture on a magnetic stirrer and leave it stirring for 24 hours.
4. Filter the mixture and stain is ready for use.

Leishman Stain is a neutral stain for blood smears which was devised by the British surgeon W. B. Leishman (1865–1926). It consists of a mixture of eosin (an acidic stain), and Methylene blue (a basic stain) in Methyl alcohol and is usually diluted and buffered during the staining procedure.

3) Stain slides with May- Grunwald stain solution for 3 minutes. (Note :Staining time may vary depending on concentration of stain, determine optimal staining time with a trial slide before proceeding.) 4) Add equal amount of distilled water, tilt to mix and stain 1 minute.

The iron staining procedure utilizes the Prussian Blue stain for ferric iron to assess bone marrow iron stores. This procedure is particularly helpful when evaluating patients with anemia, iron overload, myelodysplasia, etc.

Diff-Quik is a commercial Romanowsky stain variant used to rapidly stain and differentiate a variety of pathology specimens. It is most frequently used for blood films and cytopathological smears, including fine needle aspirates.

Romanowsky stains are neutral stains composed of a mixture of oxidized methylene blue (azure) dyes and Eosin Y. The azures are basic dyes that bind acid nuclei and result in a blue to purple color. The acid dye, eosin, is attracted to the alkaline cytoplasm, producing red coloration.

Azure (/ˈæʒər, ˈeɪʒər/ AZH-ər, AY-zhər) is the color between cyan and blue on the spectrum of visible light. The color is named after the mineral azurite. It is often described as the color of the sky on a clear day.

Eosin can be used to stain cytoplasm, red blood cells, collagen, and muscle fibers for histological examination.