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How to Work Out a Viable Cell Count

When doing research involving living animal cells, it is very important to monitor the health of the cell population. After all, it is the purpose of most cell research to determine the effect of certain treatments or microbial organisms on live cells. One can determine the overall percentage of viable cells in culture using a technique called Trypan Blue exclusion counting. Trypan Blue is a dye that is only absorbed by dead cells, so when it is added to a cell sample only the dead ones turn blue. From there it is just a matter of counting the cells and calculating the percentages.

Things You'll Need

  • Cell population
  • Light microscope
  • Trypan blue
  • Micropipetter
  • Hemacytometer
  • Cover slip
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Instructions

  1. Prepare the Cells for Viewing

    • 1

      Use a micropipetter to mix 100 microliters of the cell suspension with 100 microliters of Trypan Blue dye.

    • 2

      Add 25 microliters of the resulting mixture to each of the wells on your hemocytometer.

    • 3

      Place the cover slip over the wells of the hemocytometer.

    Perform the Cell Count

    • 4

      Place the hemocytometer on your light microscope and view at 100 times magnification.

    • 5

      Count the number of clear, living cells in a one-square millimeter space demarcated on your hemocytometer. Count the total number of cells in the same space. Divide the number of living cells by the total number of cells and multiply by 100 to determine the percentage of cells that are viable.

    • 6

      Repeat the above step for several more one-square millimeter spaces. Calculate the average viable percentage by adding up your resulting percentage calculations and dividing by the total number of calculations. This number should be a fair estimation of the total percentage of cells in the population that are viable.

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