Teach you how to count cells on a hemocytometer

Teach you how to count cells on a hemocytometer

1. Prepare cell suspension: For cells cultured in suspension, proceed directly to the following step 2 (counting and calculation process). If the object to be counted is adherent cells, the culture should first be prepared as a cell suspension as follows.
① Terminate the culture, aspirate the culture solution, and wash the culture once with PBS.
②Add 1ml of 0.25% trypsin solution to the culture flask and digest at 37℃ for 3-5min. In the meantime, keep looking under the mirror. When the cells are rounded and close to detachment, the digestion solution is discarded.
③ Add a certain amount of culture medium (if these cultured cells are no longer useful, add PBS), pipette with a pipette to make the cells detached and make a cell suspension.
2. Counting and calculation process
①Place the counting microscope coverslip in the center of the cell counting plate.
②Aspirate the cells with a glass siphon, and let the siphon flow out the suspension from the groove of the counting plate on or under the microscope cover glass until the microscope cover glass is filled with liquid.
③ Count the total number of cells in the large square at the four corners under a microscope. For the cells under the line, only the ones on the upper line and the left line are counted, and for the cell mass, count as a single cell.
④ Count the density of the cell suspension according to the following formula: cell density = (the total number of cells in 4 large cells/4) × 104 cells/ml.
2. Matters needing attention:
①Be sure to disperse into single cells, and mix the cell suspension thoroughly before sampling and counting.
②When counting under the microscope, if there is a cell cluster composed of more than 2 cells, it should be counted as a single cell. If the cell mass is >10%, the cells are not sufficiently dispersed.

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