converting oib 12 bit stacks to 8 bit tif stacks- potential problem?

Posted by spt on Jun 16, 2011; 8:46pm
URL: http://imagej.273.s1.nabble.com/converting-oib-12-bit-stacks-to-8-bit-tif-stacks-potential-problem-tp3684207.html

Hi,

We are using an Olympus Fluoview confocal to take z-stacks before
bringing them into Image J. Because we want to analyze a large area, we
have to take 2 side-by-side stacks and then use the "stitch" function in
Image J to create a montage.

 

It turns out that the staining that we are looking at is overall,
brighter on one side of the montage than the other. Although we leave
the confocal settings identical for the two photos, it seems as if the
difference between the two sides may be exaggerated in Image J; the
"seam" between the 2 is clearly visible regardless of the blending
method we try.

 

I am wondering if the fact that the confocal is taking 12 bit images,
but the tifs are imported as 8 bit images is somehow exaggerating the
contrast between them.

 

Does this make sense to anyone and is there a workaround that anyone
knows of?

 

Thanks so much,

Susan Travers

Ohio State University