Hemoglobin F cytoacid elution staining assay

The anti-alkaline and anti-acidic properties of HbF are stronger than normal hemoglobin (HbA). Therefore, after the blood is eluted by acid, the red blood cells containing HbF are not discolored due to acid resistance, and can be dyed red by eosin. Red blood cells containing HbA or only a small amount of HbF are eluted by acid, leaving only the empty shadow of the erythrocyte membrane. Basic Information Specialist classification: growth and development examination classification: blood examination Applicable gender: whether men and women apply fasting: fasting Tips: Before you check, pay attention to rest and ban diet. Normal value Coloring cells 0 to <1%. Newborn 2/3. Clinical significance (1) Most red blood cells are stained with red β-thalassemia severe type. (2) A small number of red blood cells are stained with red β-thalassemia, mild aplastic anemia, and hemolytic anemia. (3) Dyeing deep hereditary HbF hyperplasia homozygous. (4) Dyeing shallow hereditary HbF hyperplasia heterozygous subtype. (5) Dyeing unevenness β-thalassemia mild type. Precautions (1) After the blood film is made, it is stained within 2 hours, otherwise a false positive reaction may occur. The smear is required to be thin and the cells are spread and spread. (2) The pH, temperature and elution time of the buffer should be strictly controlled, otherwise the measurement results will be affected. (3) Dilute Wright's staining solution can also be used for contrast staining. Inspection process (1) The blood smear was prepared according to the conventional method, naturally dried for 1 hour, and then fixed with 80% ethanol for 5 minutes (the citrate anticoagulation was stored in a refrigerator at 4 ° C for 3 days), and washed with water to be dried. (2) The fixed blood film was placed in a phosphate buffer solution of 37 ° C, pH 3.3, and immersed for 5 minutes. (3) After rinsing and drying, the leukocytes were stained with hematoxylin staining solution for 1 min to avoid false positive red blood cells. (4) After washing with water, the red blood cells were stained with Yihong dye solution for 1 min, then washed with water, and dried for microscopic examination. (5) Under the oil microscope, the red blood cells containing HbF were stained bright red by the red dye solution, which was positive red blood cells; the cell membrane leaving only the light shadow was negative red blood cells. Count the percentage of positive red blood cells in 1000 red blood cells. Not suitable for the crowd Hemophilia and diffuse intravascular coagulation. Adverse reactions and risks Risk of infection: If you use an unclean needle, you may be at risk of infection.