Posted by
Joel Sheffield on
Apr 25, 2014; 2:59pm
URL: http://imagej.273.s1.nabble.com/Quantification-problem-more-values-despite-weaker-band-tp5007430p5007437.html
Hi Verena,
You may have the lut reversed. If you move the cursor over the bands in
the image, do you get a high or a low value for the "darker" band? It
seems that you are using the Analyze particles routine for the analysis.
You might want to try the Gel Analyzer.
Joel
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail:
[hidden email]
URL:
http://astro.temple.edu/~jbsOn Fri, Apr 25, 2014 at 10:33 AM, Rena2014 <
[hidden email]>wrote:
> Hey,
>
> I´m having problems quantifying some of my blots. The problem is as
> follows:
> I have 2 lanes on my blot - one lane showing a thick protein band, the
> second one showing a much much fainter band. Surprisingly, I always get a
> higher RawIntDen for the clearly weaker band... I really have no idea what
> the problem might be.
> After development, I save my pictures as .tif (16 bit) and change it to
> 8bit
> in ImageJ. Furthermore I convert the image so that black bands appear white
> and the background is black.
> Could it be problem that I save the pics as 16bit at the beginning?
> I would be very grateful for some helping hints! Thanks!
>
> Verena
>
>
>
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