Hemocytometer calculation tutorial

In this tutorial we go through all the calculations explained in hemocytometer calculation but with a small example for both large squares (1mm wide) and small squares (0.2mm wide). We calculate the viability, the cell density, the total number of live cells and the volume to add to reach a target density.

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I did my PhD in the Department of Chemical Engineering at Imperial College London. My research focused on mathematical modeling of the cell cycle in leukemia and involved experiments with cell lines. During that time, I had to count cells with a hemocytometer so often to track growth that I got tired and decided to build an app, HemocyTap, and share my knowledge on the topic here to help as many people as possible.

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6 comments on “Hemocytometer calculation tutorial

  1. […] can proceed with the counts in the same way as in here, but this time remember to multiply by 200 due to the initial dilution you made, and additionally […]

  2. in regards to the small cells…you said in the tutorial that you counted 5 small squares. Wouldn’t you multiply by the numver of cells you counted. the 0.000004 is for one of the small squares correct?

  3. sorry, disregard previous comment. Revised…in regards to the small squares…you said in the tutorial that you counted 5 small squares. Wouldn’t you multiply by the number of small squares you counted? the 0.000004 is for one of the small squares correct?

    • Hi LeeAnne,
      You take into account the number of squares when taking the average. So you sum the number of cells you have in total among the 5 squares (in this case, 115), you divide by the number of squares (5) and you get your average number of cells per small square. 0.000004 mL is the volume in one small square (inside the central square – see here). Since you have the average number of cells in one small square, you’re good to go!
      Hope that clarifies, let me know otherwise :)

  4. Hi where did you get 2 where you multiply the dilution factor?

    • Hi Cindy,

      If you have a 1:1 dilution (considering 1 part of original sample to 1 part of dilutant), the concentration in the original sample will be doubled compared to the one in the diluted sample – that’s why you have to multiply by 2 the value of the concentration for the diluted solution.

      Check out my longer reply in the Youtube comments here.

      Let me know if you need more help.

      Maria

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