Giemsa stain

This product is a mixture of azure pigment, eosin and methylene blue. The chemical properties of various cell components are different, and the affinity for various dyes is also different. Eosinophilic particles are basic proteins, which are combined with eosin and dyed pink; eosinophilic particles, such as nucleoprotein or lymphocyte cytoplasm, are acid substances, which are combined with basic dye methylene blue or azure and dyed purple blue; neutral particles are in isoelectric state and can be combined with eosin and methylene blue, which are light purple. All kinds of components are distinguished because of their own characteristics and the combination of different substances in jimsa dye solution to present different colors.
The Giemsa stain solution prepared by our company is made of imported Giemsa stain as raw material, which can dye the nucleus to purplish red or blue purple, and the cytoplasm to pink, presenting a clear cell chromosome image under the light microscope. It is mainly used to show the difference of the shape and size of various blood cells in blood smear, with good staining effect, strong staining power and clear staining.

Hemoglobin and eosinophils were dyed pink; nucleus, lymphocyte and basophil cytoplasm were dyed purple blue or blue; protoerythrocytes, early erythrocytes and nucleoli were dyed thick blue; middle and young erythrocytes were dyed red blue or gray red; mature erythrocytes were dyed pink.

The chemical properties of various cell components are different, and the affinity for various dyes is also different. Eosinophils are basic proteins, which are combined with eosin and dyed pink; nucleoprotein and lymphocyte cytoplasm are acid, which are combined with basic dye methylene blue or azure and dyed purple blue; neutral particles of neutral substances are in isoelectric state, which can be combined with eosin and methylene blue and dyed light purple.

[Product advantages] Stable dyeing effect and fast coloring
[Storage conditions] Room temperature and dark storage
[Shelf life] Two years
[Specification] 5ml, 50ml, 150ml, 250ml, 500ml

This product is the original solution. When using, take 1ml of jimsa concentrated solution and 10ml of phosphoric acid buffer solution to mix well, and then it can be used.
Step 1. Prepare the split phase sections according to the conventional method, dry them, drop the diluted dye solution to cover all sections, and dye them at room temperature for 2-6 minutes.
Step2. Wash the slide slowly from one end with tap water (pay attention not to wash the slice directly), and conduct microscopic inspection after drying.

1. Seed blood:
Under aseptic condition, 0.3-0.5ml heparin anticoagulant whole blood was inoculated vertically into lymphocyte culture medium with No.7 needle.
2. Culture:
The cells were cultured in 37 ℃ constant temperature incubator for 68-72 hours, and the flask was shaken once every 24 hours to make the cells fully cultured.
3. Cytological treatment:
3.1 add autumn: add 3-4 drops (about 30 μ L) of colchicine with the concentration of 40 μ g / ml to the 7-needle 2-3 hours before the completion of culture, and then put it back into the constant temperature incubator at 37 ℃ for 2-3 hours.
3.2 sample collection: after the completion of culture, pour the culture medium into the corresponding number of 15 ml centrifuge tube, 2000 R / min, centrifuge for 10 minutes, and remove the supernatant.
3.3 low permeability: add 9ml of low permeability liquid (KCl) with 37 ℃ water bath in advance to each tube. Blow 3 times for each tube, then 7 times for each tube, 28-30 minutes for water bath, and 10 times for each tube in the middle of 14-15 minutes. Formula of hypotonic solution: weigh 5.6g KCl, dissolve it with 1000ml purified water, and take a 37 ℃ water bath before use.
3.4 pre fixation: add 1ml of the fixed solution with 37 ℃ water bath ahead of schedule into each tube completed by hypotonic treatment, blow three times in each tube, and then blow seven times again. After 10 minutes of water bath, centrifuge for 10 minutes at 2000r / min to remove the supernatant. Formula of fixed solution: mix anhydrous methanol and glacial acetic acid according to the volume of 3:1. It is now prepared for use. Use 37 ℃ water bath before use.
3.5 fixation: add 9ml of fixing liquid into each tube, blow three times first, and then blow seven times, water bath at 37 ℃ for 10 minutes, centrifugation at 2000r / min for 8 minutes, and remove the supernatant. Repeat this procedure two more times.
three point six Drop: (1) add 0.5ml of fixed solution to each tube if there is more precipitation, and do not add fixed solution if there is less precipitation. After blowing gently for 5 times, leave it at room temperature for 30 minutes, and then gently blow it for 5 times before dropping and mix it; (2) take out the micro frozen glass slide, mark it, drop 3-4 drops of sample, and bake it for 1 minute at 60 ℃; 3) put the drop into 90 ℃ electric blast drying oven 1 5 hours.
4. Chromosome banding:
4.1 pancreatin digestion: put the processed slides into 0.025% pancreatin solution (ph7.2-7.4) which is preheated to 37 ℃ for digestion for 4 minutes and 20 seconds, and then rinse them in 0.9% NaCl solution which is preheated to 37 ℃ for 3 times.
4.2 dyeing: dye in Giemsa solution that is preheated to 37 ℃ for 3 minutes, wash with tap water, dry or dry, and then read the tablet.