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Quick Cell Counting Methods

Quick Cell Counting Methods - Cell Biology and Cell Culture

Quick Cell Counting Methods - Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.


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Old 07-09-2007, 12:02 PM
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Question Quick Cell Counting Methods



Hello everyone,
does anyone know an easier and quicker cell counting methods than using haemacytometer?

Counting cells is quite a difficult job when you have many cell samples to count.

The coulter, a particle counter, may be a substitute. However a coulter is not cheap and sometimes cells block the tube in the coulter. Are there any other good way to count cells?

Thank you in advance
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Old 09-05-2013, 07:28 AM
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Default Re: Quick Cell Counting Methods

Useful for to count cells a colorimetric assay using the basic azo dye Janus green. Method on MTP : cells suspension in MTP are centrifuge at 150 g for 10 min. at room temperature and remove suspension medium by vacuum aspiration and cells were fixed in 50% (v/v) ethanol for 30 min followed by vacuum aspiration of the fixative. Fixed cells were stained with 0.2% (w/v) Janus Green B in PBS for 3 min at room temperature. The stain was removed using a vacuum aspirator, and the whole plate was sequentially dipped into tap water and destained for 1 min. by gentle shaking. The remaining water was removed by shaking inverting plate on blotting paper, and stained cells were immediately lysed in 0.5 N HCl and absorbance readings taken at 595 nm using a microplate reader. Cell numbers were calculated from absorbance readings using the formula : Cell Number = (absorbance-0.00462)/0.00006926.
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Old 09-05-2013, 01:36 PM
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Default Re: Quick Cell Counting Methods

Method on tube : dispense 0.5 ml cells suspension in 3 ml tube and centrifuge at 250 g for 10 min. at room temperature. Remove supernatant medium by vacuum aspiration and cells are resuspended and fixed in 200 Ál of 50% (v/v) ethanol for 30 min., centrifuge and followed by vacuum aspiration fixative must be removed and to make sure that the remaining alcohol is completely evaporated. Dispense 100 Ál of 0.2% (w/v) Janus Green B in PBS, resuspeding cells and incubate for 3 min. at room temperature. Centrifuge as above and remove stain using a vacuum aspirator, and add 2 ml of tap water or PBS (8 g/L NaCl, 0.2 g/L KCl, 1.15 g/L Na2HPO4•2H2O, 0.2 g/L KH2PO4, adjust to pH 7.2) and destained for 1 min. by gentle shaking, rinse 3 time, following centrifugation each time. Centrifuge as above and to draw off the supernatant and the stained cells are immediately lysed in 250 Ál of 0.5 N HCl. Mix accurately and to transfer 200 Ál of it in MTP and read the Abs at 595 nm using a microplate reader. Cell numbers were calculated from absorbance readings using the formula : Cell Number = (absorbance-0.00462)/0.00006926.
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