HuibaiReagent tells you: Rayleigh dyeing process

Rayleigh staining is one of the most common and simple staining methods at present. The mixture of acidic eosin and basic methylene blue in Reese reagent was chemically treated to become neutral eosinized methylene blue. After long-term placement, azurol was oxidized to contain azure. These three dyes combined with NH3 and COO- in the nucleus and plasma, respectively, and stained the nucleus and cytoplasm. Due to the neutral dye and the buffer to regulate the acidity and alkalinity, the cells were stained with blue-red and other colors were more moderate, nuclear dye, cytoplasm and the granules in the color was more clear.

the first phase is the combination of an acid dye ihong and an alkaline substance in the cell, such as hemoglobin, eosinophilic granules, and the like; and the alkaline dye, such as a nuclear chromatin (chro-matin) and a specific neutral particle, is in contact with an acidic substance in the cell, Platelet and the cytoplasmic binding of the nuclear protein. The second phase is azure B-eosin complex, which is the basis for the formation of the Romanovsky-Giemsa effect.

The Romanowsky-Giemsa effects include(Reagent:
1. The chromatin of leukocyte nucleus was stained purple, and the chromatin of Plasmodium malaria was red.
2. the particles of the neutrophils and the platelet granule area are dyed in purple;
3. The effect of this effect must be two kinds of dye participation, one is Tianqing B and the other is Eosin.
Azure B binds to the phosphate group in the DNA molecule, and Iris binds to both the cation site in the DNA molecule and the azure B molecule. The binding power of the azure B to the azure B comes from the electron transfer power of the electron donor-receptor. A hydrogen bond is formed between the methyl amino group (- NHCHs) of azure B and the treason (- COOH) in the eosin molecule. Therefore, azurol B is the best choice for thiazide dyes to form complexes with eosin, because methylene blue with four methyl side chains cannot bind with eosin by hydrogen bonds.

Methanol, except as solvent of dye, can dissociate Rayleigh dye into azure B with positive charge and IIR with negative charge. Because of its dehydrating capacity, methanol can also fix cells in a certain form and precipitate protein into granular, reticulated structure. Increase the contact surface between cells and dyes, enhance the dyeing effect.

Various organic substances (especially proteins) and dyes in cells are sensitive to the state values of the environment. If the environment is partial acid, the hydrogen ion increases, reduces the affinity to the basic dye, enhances the eosin coloration, appears abnormal red dye, on the contrary, may appear abnormal blue dyeing. The optimum environment should be maintained at pH 64? 6, 8, and must be buffered.

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