Hi all,
I haven't found the older post mentionned, so I figured I'd just post this in reply.
Unfortunately, it is not just about not using the grid. Cell counting chambers are specifically made to count cells in a volume (As in, there is a precise separation between the coverslip and the chamber - Usually 0.1mm) which allows you to do the counting accurately, even if you pipet a semi-variable amount each time. So if you want to use automatic methods, you still need a cell counting chamber-like device. It's not enough to put liquid on a glass slide and put a coverslip on it, the volumes would not match and probably be much less accurate.
Even if you manage to get a counting chamber-like device without the grid, you still need to keep the stereology counting rules like for the manual counting (Keep objects if they intersect the bottom and left corners, discard if they are in contact with the upper and left corners) in your counting pipeline, or you rist over/under estimating the amount by a fair percentage.
All in all, you risk having a less accurate count by doing it automatically than if you counted a few chambers manually... And loose more time in the acquisition and processing then if you had just counted them by hand.
Best
Oli
Hi Tizy,
as far as I can remember something like this has been on the list a while ago - the final solution was simply to avoid the grid. You need to calibrate the scale of your image separately (with your grid or any other object of known size) and keep the magnification.
Michael
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On May 17, 2012, at 21:20, tizianarossetti wrote:
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