Thanks for the answeres so far!
I am using the Leica SP8 microscope and i'm using 8-bit images. The Problem is, that i cannot get an continous histogram in ImageJ and i am worried about losing some information. maybe it might be due to the reverse engineering of the .lif file protocol...? Furthermore, if i convert the images from RGB to 8-bit and back, it changes every time...
Thanks,
Stefan
>>> "Michael Schmid-3 [via ImageJ]" <ml-node+[hidden email]> 5/15/2015 3:23 PM >>>
On May 15, 2015, at 10:53, stefan_dillinger wrote:
> i have a bunch of -lif files and try to look at the histograms. When i look
> at the histograms directly, they seems to be binned. Only values ending on 0
> or 5 show any values.
On May 15, 2015, at 12:07, stefan_dillinger wrote:
> The problem is that the histogram in the life 8-bit image shows
> this binning and i am not sure why this is the case, it should be
> continuous, right?
Hi Stefan,
it depends on how your data were generated. E.g. if you have a sensor with 12 bits resolution, the acquisition software might multiply the raw data with some value to better use the full 16-bit range of the data and also to reduce the digitization noise upon further operations such as background subtractions and flat field correction.
For 8-bit data, this is more unusual. Some integrating cameras (Mintron, WaTeC) with analog video output seems to use only 5 or 6 bits, not 8-bit values when used with exposure times longer than the TV frame rate. If the analog gain is not exactly 1, this may result in steps of roughly 5.
If you post which instrument you have used, maybe someone knows more about it.
Michael
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