How to identify actin stain and degree of actin struktur

Hi.

Thank you for a fantastic software. I will absolutely include it in my sitations.

I am interested in calculating the degree of fluorescent stained actin structure in animal cells. Preferably I would like to identify the intensity of actin filaments throughout the cell and not the average intensity (including the not stained cell areas). I would also like to be able to calculate the degree continuous structure of actin filaments. For example 1 being prefect structure and 0 being scattered or no filament structure. Finally, if possible, it would be great to be able to measure the width of the filaments.

Is there a module or combination of modules that could solve this problem?
Just solving one of these questions would help a lot.

Best regards
Emilie

Hi Emilie,

CellProfiler doesn’t have specific modules to perform the task you’re describing. However, I can imagine that you might be able to use EnhanceEdges to enhance the filament edges, if they exist, and then detect the edges as objects using IdentifyPrimaryObjects. The difficulty is that if there are no filaments, EnhanceEdges might still find spurious edges but you may be able to set the lower threshold limit in IdentifyPrimaryObjects to get rid of them.

To assess the amount of continuous structure is more difficult. Perhaps the number of edge objects or the average length of edge objects found might give you a clue? Hopefully, contiguous filaments won’t be as numerous or as short as cells with non-contiguous filaments.

It’s difficult to assess whether CellProfiler can assist in your assay without some images to look at. Would you mind uploading a couple to this forum thread?

Regards,
-Mark