Giemsa stain

Giemsa stain is a composite dye composed of methylene azure, methylene blue, eosin and other components. Giemsa stain stains chromatin, nuclear membranes, specific cytoplasmic components and microorganisms. Giemsa stain is used in research on cytological and parasitological staining^[1][2][3]. In Vitro:Guidelines (the following is our recommended experimental protocol. This protocol is provided as a reference only, and specific procedures should be adjusted according to your actual needs).
Giemsa stain staining protocol^[4]:
1. Preparation of Giemsa stain stock solution
Dissolve 1 g of Giemsa stain powder in 66 mg glycerol and heat in a 56℃ water bath for 90-120 minutes. Subsequently, add 66 mL of anhydrous methanol and mix thoroughly. Seal the container and allow it to stand at room temperature. The solution should also be filtered before use.
2. Preparation of Giemsa stain working solution
Dilution ratio 1:10-20: Giemsa stock solution: 5-10 mL, Sorensen buffer (pH 6.8): 95-100 mL
3. Staining of blood smears
a. Fix the specimen slide in anhydrous methanol for no less than 30 seconds.
b. Tilt the slide or remove it from the fixation jar to eliminate residual methanol.
c. Prepare freshly made Staining Solution No. 1 (May-Grunwald working solution, diluted with an equal volume of buffer). Place the slide flat and immerse it in the staining solution, or place it in a staining jar for 5 minutes.
d. Without washing, directly transfer the slide into freshly prepared Staining Solution No. 2 (Giemsa stain working solution, diluted with 9 volumes of buffer). Alternatively, allow the staining solution to drain from the vertically held slide before immersion. Stain for 10-15 minutes.
e. Discard the staining solution and rinse the slide once in a buffer bath.
f. Wash the slide thoroughly with running water.
g. Immerse the slide in a clean water bath for 2-5 minutes.
h. Place the slide at an angle and allow it to air dry naturally. Do not blot dry with absorbent paper.
i. Mount with a coverslip if required.