Posted by
Krs5 on
Oct 31, 2014; 11:22am
URL: http://imagej.273.s1.nabble.com/Help-needed-with-method-of-background-substraction-tp5010244p5010263.html
Dear Abs,
Try if when you select under Analyze > Set Measurements the option 'Limit to threshold' your measurements are as expected.
Best wishes
Kees
Dr Ir K.R. Straatman
Senior Experimental Officer
Advanced Imaging Facility
University of Leicester
http://www2.le.ac.uk/colleges/medbiopsych/facilities-and-services/cbs/lite/aifImageJ workhops 15 and 16 December
-----Original Message-----
From: ImageJ Interest Group [mailto:
[hidden email]] On Behalf Of khanas
Sent: 30 October 2014 15:32
To:
[hidden email]
Subject: Help needed with method of background substraction
Hi all,
Have come across an image processing issue this morning and would really appreciate your help! Forgive my ignorance please I am quite new to microscopy/imageJ!
Essentially to quantify nanoparticle uptake into cells we take images with a whole cell fluorescent stain and a reflectance channel - the nanoparticles turn up in the reflectance channel and we use the staining to draw around a cell and analyse it for mean intensity.
To remove background reflectance other people in my lab have first split the channels of the image, and then using the max and min controls in B&C changed the min value until any background has been removed. After this they have selected the area in question (using the aforementioned stain), and then Measured mean intensity.
Unfortunately for some reason, potentially due to an update, it now doesn't measure between the values we set. it only ever gives mean intensity of the unedited/adulterated image and I have verified this by measuring the same area a few times with different Max and Min settings.
If I save the file as another format this can work but such a compression would be a problem from a quantification point of view!
A colleague just showed me this forum and I will have a look through to see if there are any existing solutions but any and all help would be very much appreciated!
Many thanks
Abs
Any help would be very much appreciated!
Thanks
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