Posted by
G. Esteban Fernandez on
Apr 25, 2014; 3:49pm
URL: http://imagej.273.s1.nabble.com/Quantification-problem-more-values-despite-weaker-band-tp5007430p5007434.html
Even if the bands appear white they could still have a lower intensity that
the black background. This happens if you inverted the display colors but
not really changed the actual intensity values; the image window may say
"Inverting LUT' on the info bar.
First try applying Image > Lookup Tables > Grays to see the "true"
intensities (high intensity as white, low as black).
To confirm the intensity values, wave the mouse over a band and over
background areas whilst checking the intensity values reported in the
ImageJ window. If the bands are lower intensity then you have to invert
your image again, or don't invert it in the first place.
Good luck!
-Esteban
On Fri, Apr 25, 2014 at 7:33 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
>
>
>
> --
> View this message in context:
>
http://imagej.1557.x6.nabble.com/Quantification-problem-more-values-despite-weaker-band-tp5007430.html> Sent from the ImageJ mailing list archive at Nabble.com.
>
> --
> ImageJ mailing list:
http://imagej.nih.gov/ij/list.html>
--
ImageJ mailing list:
http://imagej.nih.gov/ij/list.html