Stevej:Cell counting/hemocytometer

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(Procedure)
(Procedure)
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#Combine 20μl cell suspension with 20μl of Trypan Blue. Mix.
#Combine 20μl cell suspension with 20μl of Trypan Blue. Mix.
#Load 10μl into each of the 2 hemocytometer "sides" using the capillary action created by the coverslip and the hemocytometer. Avoid bubbles.
#Load 10μl into each of the 2 hemocytometer "sides" using the capillary action created by the coverslip and the hemocytometer. Avoid bubbles.
-
#Count the number of cells in 4 corners of the gridded area of each side. (See below; Blue circles) White cells are live and blue cells are dead/dying.
+
#Count the number of cells in 4 corners of the gridded area of each side. (See below) White cells are live and blue cells are dead/dying.
#Record cell count and repeat for the other side of the hemocytometer.
#Record cell count and repeat for the other side of the hemocytometer.
#If the two cell counts are roughly equivalent, average the cell counts. Otherwise, recount.
#If the two cell counts are roughly equivalent, average the cell counts. Otherwise, recount.

Revision as of 13:05, 28 August 2007

Contents

Overview

Counting cells with a hemocytometer.

Materials

  • Tube of cell suspension
  • Hemocytometer
  • Trypan Blue 0.4%


Procedure

  1. Combine 20μl cell suspension with 20μl of Trypan Blue. Mix.
  2. Load 10μl into each of the 2 hemocytometer "sides" using the capillary action created by the coverslip and the hemocytometer. Avoid bubbles.
  3. Count the number of cells in 4 corners of the gridded area of each side. (See below) White cells are live and blue cells are dead/dying.
  4. Record cell count and repeat for the other side of the hemocytometer.
  5. If the two cell counts are roughly equivalent, average the cell counts. Otherwise, recount.
  6. Mean cell count/4 * Dilution = Cells x 104/ml.
  7. Above example, Mean cell count/4 * 2 = Cells x 104/ml.
demo

Notes

References


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