Re: lamp temperature
Posted by
Nathaniel Ryckman on
Jun 02, 2011; 6:43pm
URL: http://imagej.273.s1.nabble.com/lamp-temperature-tp3684358p3684361.html
Kathleen McMillan wrote
Hello- I am looking for advice on whether I can correct microscope images for variations in lamp filament temperature when analyzing slides for stain intensity. In a first batch of slides, the background was white, the (unstained) tissue was tan and the stain blue. Using the Landini thresholding plugin to select for blue hues followed by measuring the mean saturation worked well. However, in a second batch of slides - which were intended to be combined with the first in the analysis - the slide background and the unstained tissue are a yellowish color and the stain hues are in the blue-magenta region. That creates a problem in separating the tissue from the background, and also the mean saturation of the hues attributable to the stain are different from the first batch. Is there a good way to correct for lighting variation in these two image sets, that would allow an accurate and consistent stain intensity to be quantified?
Thanks for any advice!
That sounds like a really tricky problem. I highly doubt that there is an easy solution.
The only things that I can think of is perhaps somehow normalizing colors across images by creating a custom plugin:
http://imagej.588099.n2.nabble.com/help-on-normalizing-background-across-pictures-td4893699.htmlIf you think it's because of uneven lighting, perhaps, you could split the color channels, perform intensity corrections (using rolling ball/flatfield), and then merge them back together:
http://www.macbiophotonics.ca/imagej/image_intensity_proce.htmOne last idea is that you could increase the threshold range to pick up colors from blue to blue-magenta (aka not be as selective about the regions that are automatically chosen).