Chloroacetate AS-D naphythol AS-D chloroacetate esterase (AS-DNCE) staining solution (chemical coloring process)
A technology of AS-D and chlorinated acetic acid, which is applied in the field of medical in vitro diagnosis, can solve the problems of unfavorable observation of granulocyte line positive cells, affecting the diagnosis and typing of leukemia diseases, and smears cannot be stored for a long time, and the dyeing effect is not easy to decolorize. , The dyeing effect is stable and the dyeing effect is lasting
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Embodiment 1
[0041] Embodiment 1: the composition and preparation method of kit of the present invention
[0042] Fixative solution: take 10mL of 40% formaldehyde, weigh disodium hydrogen phosphate (Na 2 HPO 4 ) 0.65g, sodium dihydrogen phosphate (NaH 2 PO 4 ) 0.4g, add 90mL of distilled water, the pH value is 7.3, pack in 2.5mL per bottle, seal, and store in a refrigerator at 4°C.
[0043] Incubation Solution A (AS-D Naphthol Sodium Chloroacetate Solution): Weigh 50mg AS-D Naphthol Sodium Chloroacetate and dissolve it in 5mL N,N-Dimethylformamide (DMF). pack, seal.
[0044] Incubation solution B (diazo agent-fast red B): weigh 30mg fast red B powder and put it in a reagent bottle.
[0045] Incubation solution C (buffer solution containing saponin): weigh disodium hydrogen phosphate (Na 2 HPO 4 ) 8.8g, potassium dihydrogen phosphate (KH 2 PO 4 ) 0.9g, saponin (Saponin, purchased from Sigma) 0.1g, sodium azide 1g, dissolved in 1000mL of distilled water, pH value is 7.6, divided int...
Embodiment 2
[0047] Embodiment 2: the staining method of kit of the present invention
[0048] 1. Prefabricated dyeing incubation solution: take 1 bottle of incubation solution A, incubation solution B and incubation solution C prepared in Example 1, mix them, shake gently and mix well, and then make the dyeing incubation solution;
[0049] 2. Add the fixative prepared in Example 1 pre-cooled at 4°C to the fresh bone marrow cell smear or blood cell smear dropwise to cover the smear, rinse with water 3 times after 30 seconds, and dry the smear naturally;
[0050] 3. Pour the dyeing incubation solution in step 1 into the dyeing tank, place the smear in the dyeing incubation solution, and place it at 37°C for 30 minutes, then rinse the tank with running water for several minutes, and dry the smear naturally;
[0051] 4. Add the counterstain solution prepared in Example 1 dropwise to the dried smear to cover the smear, rinse with water for 3 times after 1-2 minutes, dry the smear naturally, an...
Embodiment 3
[0052] Embodiment 3: normal human blood cell smear detection
[0053] Get normal person's peripheral blood to make blood smear, use kit of the present invention to carry out staining to blood smear according to the method for embodiment 2, blood smear after dyeing observe blood image and take pictures with microscope oil lens, as figure 1 .
[0054] Judgment of staining results: Negative result is colorless cytoplasm; positive result is red precipitate in cytoplasm: cytoplasm is light red (+), bright red precipitate is full of cytoplasm (++), dark red precipitate is full of cytoplasm ( +++).
[0055] Dyeing result: by figure 1 The results showed that normal mature neutrophils were positive, and monocytes were negative.
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