Blood count analysis can be used to (1) get basic information about the patient's blood and (2) find out if the patient has an infection, anemia, or coagulation disorders. It is generally a mandatory test.

Operation method

blood count

Principle

The hemocytometer contains 2 chambers, each of which has a total volume of 9x10-3 ml when filled and covered with a coverslip, and each chamber has 9 large squares, so each large square has a volume of 0.1 mm3 or 1.0x10-4 ml when covered with a coverslip, and for counting purposes, the segregation of the 9 large squares is unnecessary and can be ignored.

Materials and Instruments

Cell Samples
Trypsin Cell culture medium
Capillary pipette Slide Coverslip

Move

1. Treat the cells with trypsin as described in the previous protocol (see "Trypsin Treatment of Isolated Cells") and resuspend the cells in cell culture medium.

2. Pipette the two cell specimens to be counted using a capillary pipette. The cell suspension is fed into the coverslip-covered counter by the action of the capillary pipette.

3. Count the cells in 5 of the 9 large squares on each side of the cell counter, i.e., 10 squares in total.

4. Combine the cell counts of the 10 large squares (5 per chamber, 2 chambers in total), and calculate the cell counts in ^1x10-3 ml ( ^1x10-4 ml per large squarex10 large ^squares=10-3 ml volume). Multiply the total number of cells by 1000 to obtain the number of cells per ml of counted cell suspension.

5. Immediately after counting, wash the counter and coverslip with double-distilled water, then with 70% ethanol, and wipe dry with microscope paper.

Caveat

Reasons for certain errors in applying counter counts:- Uneven sampling from the original cell suspension. The original cell suspension should be stirred and mixed well, and the cells should not settle to the bottom of the container during cell counting.- Insufficient filling or overfilling of the counting chamber. The volume of the counting chamber is based on a coverslip placed smoothly over the edge of the counter; overfilling increases the volume being counted.- Clusters of cells do not readily enter the counting chamber by gross suction, so small clusters of cells that are excluded from counting and enter the counting chamber are more difficult to count accurately. A well-dispersed cell suspension is essential for accurate counting, and the cells should be thoroughly mixed to achieve homogenization.

Common Problems

Sourced from Guide to Cellular Experimentation (Previous Volume) by Peitang Huang.

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