Posted by
Alan Thames on
URL: http://imagej.273.s1.nabble.com/Help-colony-counting-with-image-J-tp5004887.html
Dear imageJ users,
I have an issue with cell colonies
counting
using image J for my drug treated cells in 3 D soft agar. My control( non
treated) images
have big colonies in which cells are fused and in the drug treated images,
sizes of the colonies are reduced (with increasing dosage of drug) . As
the size of colonies between no treatment and drug treatment is much
different, i cannot get a proportionate decrease in colony numbers as
i can see with average colony "size". For example; in no treatment picture
- colony number may be less but it's very obvious there r huge colonies,
using the same setting for treatment given pics - colony count may be
increased due to tiny small colonies though big colonies are not present.
It makes me confused that the drug doesn't reduce colony count, but the
size.
Can any one help me how to get a setting that could count numbers
proportionate to treatment efficacy and at the same time it would detect
the decrease in size?
If i increase the size setting just
to count big colonies, the small ones are all missed. If i decrease the
size to include all colonies including tiny ones, the count is still
increased in cells treated with higher drug concentrations, due to the
presence of
tiny smaller colonies, even the big ones are not seen.
My steps are imageJ --> Type--> change the image to 8 bit format
-->adjust--> threshold--> set ; lower
threshold - 0, upper threshold- 155
--> Analyze --> analyze particles
-->size-->25 to infinity, cicularity 0.25 to 1, show outlines, Check
-display results, clear results, summarize, exclude edges
Regards,
Alan
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