I would like to perform some signal intensity quantifications on orthogonal views and I am trying to understand the image quality of these views.
I have stack images at 100x magnification of stained cells with 2 channels (dapi and RFP). The images I´ve taken are 16-bit.
I noticed that if I open the stack as a composite image and then create the orthogonal views, the orthogonal views are RGB images that upon splitting channels are 8-bit. Is there a way to preserve the bit depth in the orthogonal views? My understanding is that its not the best choice to perform the quantification on the 8-bit because I could be losing information.
I understand that I can open the stack as either colorized image or grayscale image and then indeed I see that the orthogonal views are also 16-bit. However, in these, I have my 2 channels separately so if I wanted to make a nice image, with pseudocolors and with both channels in one (say for a publication or presentation), I would need to either merge the two or open the image as a composite and somehow find the exact same position for the orthogonal views.
Essentially - is it possible to generate orthogonal views from a 16-bit, composite stack image that are also 16 bit?
I´m sorry if this is a stupid question. This is my first post here.