The BEST cell counting technique? - (Dec/13/2007 )
General question... what do you think is the best (i.e. most accurate and easiest to perform) cell counting technique? e.g. Hemocytometry, WST-1 assay, FACS, coulter counter, other??? Thanks.
i'd say the MTT assay is quick and accurate. anything where you just add solution to the well, and then measure it, is sweet bliss. alas, i use the coulter counter. accurate (because i'm the one who calibrates and cleans it AFTER EVERYONE, *tick tick*. no i'm not bitter), but time consuming. but a lot faster that the heamocytometer.
The Haemocytometer has the advantage on working on a small sample of living cells, though. To do an MTT assay one has to grow the cells separately and then kill them to get data. MTT is great for measuring proliferation, though.
Ok, I'm not somebody who usually promotes a company, but for cell counting we use our Guava system and it's the BEST!!!
Also because we usually don't count 1 sample (although you can do that to) because it has a 96 well format.
Basically you can stain your cells with live/dead dye like 7aad or use their solution (of course this is more expensive, but you don't have to wait to stain. You just add it, put it in the machine and you can walk away.. ok, after you turned the run on!)
It's like a small FACS machine. You have forward and side scatter and you have 3 channels (I think the newer ones have even more) so also if your FACS machine is not working, you can use it. Or if you have so many samples, but not a lot of colors, you can use it. And the best thing is. You put it in the plate, put in the format which wells you want to count, hit start, walk away and look at it when it's done!!.. Which is the best esp. having a lot of samples and other stuff that you can do in stead of sitting next to the FACS machine and wait to put the next tube in...
Ok.. I really love it. sorry
i would say... depends on your budget and the sample size you work with.
If you just want to count cells once in a while when subculturing... hemacytometer is the best. For MTT, you'll need certain minimum number of cells to get the accuracy. too much/less cells won't work. and also the incubation time before detecting... needs couple of hours, Hemacytometer is straight forward. My friend actually just using hemacytometer to do her cell viability curve, counting with this method for every 2 hrs time-point. And it actually produce a nice graph!!! who says cheap method won't work? However, i would prefer MTT if I am doing what she did --- coz too many samples, too many time-point and she count like crazy.
If you have budget to go for something automated... then, no harm to try right?
can't say which one is better. depends on your usage and MONEY, of course.