Count cells without Dapi

Hi,

I hope you can help me. I want to count cells in each picture and I know that I should use IdentifyPrimary object and then IdentifySecondary object but the problem is that my DAPI staining failed so I only have a good phalloidin signal. Is it possible to identify cells without the use of IdentifyPrimary objects? I have nice single cells so I guess it should be possible to identify the each cell with just the phalloidin signal.

Thank you.

Marloes

Hi Marloes,

Could you mind posting a few example images, so we can take a look?

Thanks!
-Mark

Hi Mark,

Here are 2 images from the txred (phalloidin) channel.

Marloes

Hi Marloes,

Unfortunately, based on these images, identifying the single cells will not be possible. CellProfiler typically relies on intensity gradations to determine cell boundaries, and if the cells were well-separated, it could still work without a nuclei stain. In other cases, the cell stain sometimes has a dark “hole” where the nucleus would be that could be used as a surrogate for the nucleus itself. However, neither of these situations are true in your case.

I should mention that even if the DNA stain had worked, CellProfiler would have a tough time figuring out the cell boundaries nonetheless, since the cytoskeleton doesn’t have much in the way of intensity gradations that permit separation of overlapping cells.

Regards,
-Mark

Hi Mark,

Thanks for you reply.

Marloes