Eosinophilic spot-colored erythrocyte counting assay

Summary

Dot-colored red blood cells is a kind of more naive red blood cells, the cells contain hemoglobin, but also contains basophilic substances, the alkaline dye stained to form a large number of small blue particles, known as dot-colored red blood cells, unlike the reticulocytes, dot-colored red blood cells are used for drying blood membrane staining display, because the dot-color is formed during the process of cell drying.

Operation method

Eosinophilic spot-colored erythrocyte counting assay

Principle

The basophilic material in the red blood cells is shown as blue dots in the red blood cells using Rachel's or methylene blue stains, and the dotted red blood cells are then counted as a percentage of the total number of red blood cells.

Materials and Instruments

Blood
Alkaline methylene blue solution Sodium bicarbonate Ricci's stain

Move

I. Experimental reagents:

1、Basic Meilan solution Meilan 0.5g sodium bicarbonate 3.0g, distilled water Add to 1000ml

Dissolve and filter with filter paper, this reagent can be used for 2-3 weeks, if there is precipitation should be discarded, re-formulation.

2、Richter's dye: Richter's dye can show point color red blood cells, but not as clear as alkaline dyes, so more often use the American blue staining method.

Second, the experimental operation:

1, with the usual method to make a thin blood film, dry and fixed with methanol, 2, with alkaline Meiyi blue dye 40s water washing, drying.

3、Use low magnification to select the part of red blood cell with uniform distribution, and then change the oil color counting, the red blood cell is strong blue, point color particles are dark blue.4、Counting method: basically the same as reticulocytes, counting the point-color red blood cells in the field of view of 50 at the same time counting the total number of red blood cells in the field of view of 5 of them, you can calculate the percentage of the point-color red blood cells For example; the point-shadowed red blood cells in the field of view of 50 is 15, and the total number of red blood cells in the 5 The total number of red blood cells in the field of view is 1000, then the spot color red blood cells = (15/100 × 10) × 10% = 0.15% 5, such as the use of Rachel's staining method with the blood film staining

Caveat

1, the blood film should be spread thin. So that the red blood cells do not overlap.

2， the drying speed should be slower after the blood film is made, so that the point-shaped particles formed are larger.Reference value: normal ＜0.3%Significance: Heavy metal (lead, bismuth, silver, mercury, etc.) poisoning and nitrobenzene, aniline and other poisoning, spot color red blood cells increased significantly, hemolytic anemia, pernicious anemia, iron granulocyte anemia, white blood and malignant tumors, spot color red blood cell percentage is also higher.

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