Synchronizing and Combining Tiffs


I am trying to synchronize and combine two separate tiff stacks that are the same size, depth, and type. I was able to sync them (Fig 1), but when combining them into the same file, I always lose one or the other (Fig 2 & 3) in quality or visibility. Does anyone know why this may be happening? Thanks in advance.




Check whether the greyvalue ranges for both stacks are in a similar range (while they are both the same 16-bit data type, maybe these are coming from different cameras with very different greyvalue ranges e.g. 10bit vs. 14bit.).
Have you played with contrast/brightness of your combined stacks?

You are right, they do seem to have different greyvalue ranges (one is 0-255, and one is 3-65535). Thank you for pointing this out. Will changing the contrast/brightness fix this? I don’t want to lose the brightness integrity of the calcium video since I need to see how the cells fluoresce.

Also, when I transferred the files to my home computer, a Mac (ImageJ v1.52k), I was able to do it without any problems, which is quite strange.

Well, if you multiply the 8 bit image by 256 you will bring it in the same range as the other one.
This doesn’t lose any information, but it will create gaps in your histogram which could potentially upset some analysis algorithms. But running any sort of analysis on the fused image does not make sense anyway IMHO.
I have no explanation for the difference in behaviour you see on your Mac.