There are a number of other methods for counting cells, most of which require expensive equipment such as: electrical impedance, image recognition, etc. However, if following a 1:10 dilution you are only averaging 5 cells per 0.1mm3 I would suggest skipping the dilution protocol completely. If you avoid dilution then you would still only predict to need to count ~50 cells per 0.1mm3, which isn't as time consuming as multiple pipetting steps. Not to mention it minimizes the human error factor of pipetting. Hope this helps.

Now do you have a different cell population for each of the 96 wells? If that is the case I would suggest looking into an automated counter.

Our laboratory uses an automated counter (Coulter Counter) to save time on counting cells. This is much more convenient and saves a lot of time.

We use CASY cell counter. You just need to dilute your cells (1:100, 1:400) in the Casy buffer and measure taking into account the settings for your cells (in which size range they are). The advantage is that you can measure cell viability, which is determined by electric current exclusion, so you don't need viability dies. Also, it is more accurate that the Coulter counting, since the technology used is not based in an electrolytic reservoir, where the cells are counted batch by bactch and any particle able to displace the electrolyte would be counted.

If you can use a flow cytometer cell sorter you can deposit exactly 3000 (vital that means by example PI neg.) per well.

As already suggested a flow cytometer might work if the cells are uniform in shape. I am familiar with an instrument by BD which allows you to count and sort populations into tube for other downstream applications, i.e. cultivation or single cell genomics.

With FACS ARIA from BD or MoFlo from Beckman you can deposit single cell or another count of cells in any well plate you want.

Dear Bidisha,

I agree with all the above answers. However, I just want to add a little correction to your calculation. You have to multiply by the dilution factor, 10 in your case.

Regarding a faster counting tool, I would suggest Scepter Handheld Automated Cell Counter from Millipore. Here is the link to it:

http://www.millipore.com/userguides/tech1/00110230tp1

I hope this helps.

Dr. Abdelkader Ashour

I love the scepter. Best money I have ever spent on a small piece of equipment.

We use Countess® Automated Cell Counter from life technologies! It is really recommendable.

Counting cells when they are growing adhesive is not that easy. But there is a simple trick: each cell contains a constant content of a protective enyzme, the Cu/Zn SOD.

You need to run only once a calibration using a defined number of cells lysed by freezing/thawing or by Tween 20 or Triton X100. After centrifugation of the cell culture plates and removal of most of the cell culture media you can lyse the cells (as described above: Triton X100 is the best way) and measure the duluted lysates in the ELISA in the link below. The owner of this company can give you some further suggestions.

It's simple and easy to do.

http://www.seramun.com/products/elisa-kits/human-proteins-and-antibodies.html