Stevej:Cell counting/hemocytometer: Difference between revisions
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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 the gridded area of each side. (See below) 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; Blue circles) 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 10:49, 28 August 2007
Overview
Counting cells with a hemocytometer.
Materials
- Tube of cell suspension
- Hemocytometer
- Trypan Blue 0.4%
Procedure
- 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.
- 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.
- 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.
- Mean cell count/4 * Dilution = Cells x 104/ml.
- Above example, Mean cell count/4 * 2 = Cells x 104/ml.
Notes
References
