White Blood Cells (WBC), formerly known as leucocytes, are the "guards" in the human body’s fight against diseases and a vitally important group of immune cells in human blood.
WBC dilution, all mature red blood cells are lysed. The mixture is then loaded into a hemocytometer, and the number of white blood cells in a defined volume is counted under a microscope. The white blood cell concentration per liter of blood is subsequently calculated via conversion.
Functions:
1. Dilute white blood cells in the blood to prevent the aggregation and adhesion of blood cells.
2. Maintain stable physicochemical parameters such as pH, conductivity and osmotic pressure.(Note: The set pulse signal thresholds vary among different instruments when measuring samples via the electrical resistance method.)
This WBC diluent is for research use only to facilitate white blood cell counting, and not intended for clinical diagnosis.
Operating Procedures (For Reference Only):
1. Add 0.38 mL of WBC Diluent to a small test tube.
2. Use a clean, dry micropipette to draw 20 μL of peripheral blood. Wipe off the residual blood on the outside of the pipette, insert the pipette to the bottom of the WBC Diluent, release the blood gently, then aspirate the supernatant to rinse the pipette 2–3 times and mix the solution thoroughly immediately. The standard dilution ratio is 1:20, and dilutions at 1:10 or 1:40 are also acceptable.
3. After complete red blood cell lysis (the solution turns brown), mix the solution again and load it into the hemocytometer, taking care to avoid bubble formation or overflow. Let it stand at room temperature for 2–3 minutes to allow white blood cells to settle.
4. Place the hemocytometer under the low-power microscope and count the number of white blood cells in the four large corner squares one by one. For cells overlapping the grid lines, follow the counting principle of "count cells touching the top and left lines, not the bottom and right lines".
Precautions:
1. Do not over-squeeze the puncture site during blood collection, and ensure the needle puncture depth is appropriate.
2. Keep the small test tubes and hemocytometers thoroughly clean to prevent misidentification of foreign particles as white blood cells.
3. Within the reference range, the difference in the number of cells between large squares should not exceed 8, and the difference between two replicate counts should not be more than 10%.
4. If the white blood cell count is excessively high, increase the dilution ratio; if the white blood cell count is extremely low, count the cells in 8 large squares or collect a larger volume of blood for testing.
5. For your safety and health, wear a lab coat and disposable gloves during all operations.
Calculation:
WBC count/L = (N÷4)×10×20×10⁶=(N÷20)×10⁹
N
| Total number of white blood cells in 4 large squares
|
÷4
| Average number of white blood cells per large square (0.1 μL)
|
×10
| Convert the volume of 1 large square (0.1 μL) to 1.0 μL
|
×20
| Blood dilution ratio
|
×10⁶
| Convert from 1 μL to 1 L |
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