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When to Change

Use this page when counting cells. Cell counting should be done before seeding a transwell or when passaging/splitting cells.

Purpose

It is important to know how many cells you have so that when you seed a transwell or a flask, you know how much volume of DMEM you should use to seed with when resuspending.

Safety

• Complete work in the BSC.
• Spray down all materials brought into the BSC with 70% Ethanol.
• Try not to touch the sides of the flask opening with pipette tips.
• Do not touch the rims of the flasks or tubes to work with.

Materials

• Cell culture flask
• Trypan Blue
• Cell Counting Machine
• 1.5 mL tube

Procedure

*Complete the adhesion removal and suspension steps of Caco-2 Splitting Cells

1. Add 10μL of cells and 10μL of trypan blue to 1.5mL tube.
2. Mix the cells and dye together by aspirating and releasing in a pipette. Add the solution to side A and B of the measurement slide.
3. Insert into the machine with the arrow of the letter to be measured facing in the direction of the insertion clip. Press the “count live cells” button.
4. Cells will be counted in 10^6 cells/mL. Note down the number and % of both dead and alive cells.

Quality Control

• Make sure that the trypan blue-cells solution is evenly spread out in the well of the cell counting machine slide.

Revision History