Stevej:Cell counting/hemocytometer
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(New page: ==Overview== Counting cells with a hemocytometer. ==Materials== *Tube of cell suspension *Hemocytometer *Trypan Blue 0.4% ==Procedure== #Combine 20 ul cell suspension with 20ul of Tryp...) |
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==Procedure== | ==Procedure== | ||
| - | #Combine 20 | + | #Combine 20μl cell suspension with 20μl of Trypan Blue. Mix. |
| - | #Load | + | #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 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. | ||
| - | #Mean cell count/4 * Dilution = Cells x | + | #Mean cell count/4 * Dilution = Cells x 10<sup>4/ml. |
| - | #Above example, Mean cell count/4 * 2 = Cells x | + | #Above example, Mean cell count/4 * 2 = Cells x 10<sup>4/ml. |
==Notes== | ==Notes== | ||
Revision as of 13:40, 28 August 2007
Contents |
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 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.
- 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
