Identify cells/2nd objects using 2 markers?

Hi -

Is it possible to identify 2nd objects using 2 markers? I have Phalloidin channel which is sometimes weak, and another channel that sometimes works pretty well. Occationally they stain whole cell mutually exclusively good - if you know what I mean. I assumed there is no option of using more than 1 marker in identifying objects module. Hence, I try to manipulate the masks instead. Two of 2nd objects - say phalloidin and, say, m2 are saved. From my observation, a simple union operation of phalloidin and, say 2nd marker, m2 might just do the work. I am wondering if there is a way to do so, and then extract features from that manipulated mask instead. Otherwise, I can process this outside cellprofiler, then I am wondering if there is a way to import the mask back.

Any suggestion would be greatly appreciated. Thank you for such an awesome tool and great forum.

You could try ImageMath, and add the two channels together.

Did you mean adding 2 channels before identifying the secondary objects? One channel is much brighter than the other. I guess I just need to find a right factor to mix them both. That might work. Thank you!

Indeed ImageMath is what I would suggest also. You can multiply the images before the operation within ImageMath. You could Add, or once the images are on the same intensity scale, you could try “Maximum” too.

Other operations like Morph or EnhanceOrSuppressObjects can also tend to equalize images’ (or the objects within) intensities, but I like ImageMath since it is straightforward.