Knowledge of the principles and techniques of basophil counting by neutral red method

Operation method

Direct Basophil Counting Assay by Neutral Red Assay

Principle

Dilute the blood by suction neutral red dilution to a certain number of times． At the same time, destroy the red blood cells and make the basophils stain brick red. Then drop into a counting chamber and count the number of basophils within a certain range to find the number of basophils per liter of blood.

Materials and Instruments

Neutral Red Dye Saponin Formaldehyde

Move

III. Experimental reagents:

l. Storage solution

①, PH5.4 0.1mol/L

②, 0.12g/L neutral red dye solution

③, 4Og saponin

④, 40% formaldehyde

2, working fluid:

① neutral red saponin solution in 1mL of newly filtered neutral red dye, add 40g/L saponin solution 0.25mL

② formaldehyde fixation solution: in 0.5mL formaldehyde plus phosphate buffer 3.5ml.

Fourth, the experimental operation:

1, in a small test tube add neutral red saponin solution 160ul capillary blood 20ul mixing. Add formaldehyde fixation solution 20ul, so that the blood diluted 10 times.

2, will be mixed suspension drops into the counting plate of the two counting chambers, in order to prevent evaporation should be placed in the wet combined static 1min, so that the basophils sinking coloring 3, in the low magnification microscope to count two counting chambers all the large square in the number of basophils V. Calculation:

Basophil number/L = (number of basophils in 18 large squares/18)×10×10×106 Normal reference value: 0.02-0. ^05×109/L

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