Giemsa Stain (Blood Cell/Plasmodium)

Assay-kits

Giemsa Stain (Blood Cell/Plasmodium)

Product code: ACMBS-13
Specifications: 1x100mL, 4x250mL (Box)
Intended use: This kit is for blood cell and plasmodium staining
Store temperature: 5℃～30℃
Expiry date: 24 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

Giemsa stain contains methylene blue and eosin, the former is alkaline, the latter is acidic, they have a different affinity with the various substances in the cell, so that it can show different colors to identify.

Main components

Reagent composition	Main ingredient
1. Giemsa stain (solution A)	Giemsa dye
2. Phosphate buffer (solution B)	Phosphate

Sample requirement

1. Blood cells: fresh whole blood or EDTA·K2 anticoagulant.
2. Plasmodium: earlobe or fingertip blood for thin blood or thick blood smear.

Test procedure

Fix the specimen:

1. Staining of blood cells: According to the conventional push-plate method to prepare the "tongue-shaped" blood film of uniform thickness, dried naturally.
2. Staining of Plasmodium

① Preparation of thick blood film: Take 4～5μl of whole blood (equivalent to the size of a match head) and drop it on a position about 1cm near the frosting head of the slide. Use the corner of the pusher to transfer the blood from the inside to the outside to a diameter of about 0.8～1cm uniform round shape, natural drying for more than 6 hours. If it is stained on the same day, the thick blood film does not need to be hemolyzed and stained directly, if it is left overnight, a few drops of distilled water need to be added for hemolysis for about 5 minutes, the blood is poured out, and the stain is awaited.

② Preparation of thin blood film: Take 1～1.5μl of whole blood (equivalent to 1/4 match head size) and prepare it into 2.5cm (length) × 2cm (width) uniform thickness of "tongue-shaped" blood according to the conventional push method Membrane, naturally dry.

Staining:

1. Fix the thin blood film or blood cell smear with methanol for 1 to 3 minutes and let it dry naturally, the thick blood film used for the staining of plasmodium cannot be fixed.
2. The fixed specimen is placed in Giemsa working solution [solution A: solution B = 1:10] for 10～30 minutes (if specimens were less it could be stained with drops). When the temperature is low in winter, it can stain in 37℃ incubator.
3. After dip stained, the specimens were taken out and washed with water, If using drip staining, the stain solution cannot be poured out before washing, and it should be washed away with running water to prevent sediment from settling on the specimen. Make a microscope examination after drying.

Result interpretation

1. Blood cells: pale pink erythrocytes, leukocytes nucleus stained with blue to dark blue, clear nuclear chromatin structure, clear cytoplasm particles, show a variety of cell-specific colors, such as neutrophil granule is purple, eosinophil granule is red, basophil granule is dark purple.
2. Plasmodium: Plasmodium cytoplasm is blue, the nucleus is red or purple.

Method limitation

Only used for staining of initial morphological examination and observation.

Precautions

1. Usually thick and thin blood film is coated on a glass slide, in order to prevent the blood film from falling off and affect the detection, it is recommended that outpatients and fever patients apply 2 thick blood film and 1 thin blood film.
2. To make thick blood film, pay attention to blood volume and degree of thickness. If the blood film is too thick, it will easily fall off, and if it is too thin, it will not meet the detection rate requirements, if the thick blood film is not dry enough, it will easily fall off.
3. The thick blood film should be fully dried naturally for more than 6 hours and stained within 48 hours. Plasma placed for too long will lead to poor hemolysis, and cell overlap will affect the observation of malaria parasites.
4. The staining time depends on the type of specimen, the thickness of the smear, the number of nucleated cells, and the temperature.
5. If using drip staining method, the stain solution can't be poured out before washing, and it should be washed away with running water to prevent sediment from settling on the specimen.
6. The staining solution should be shaken well before preparing Giemsa working solution to ensure the uniformity of staining. Staining solution volume should be sufficient, and do not make the staining solution evaporation to prevent the dye settling in the smear.
7. Giemsa working solution should be used immediately with the best effect within 30 minutes, the working solution is disposable, with repeated staining, the staining effect will gradually decline, and the sediment increased would lead to increase the risk of attachment.
8. This product should be used by professionals and interpretation of results.
9. Read the instructions before use, use within the validity period, and keep a personal hygiene protection.
10. After use, the disposal of waste should according to hospital or environmental protection department requirements.
11. The production date, production lot number and expiry date are shown in the outer package.

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