Licor Odyssey Western Blots

[Sieber, Anja]

Hi everyone,
I have western blot pictures, either as a two channel picture or two separate pictures of the two channels (700/800nm). I want to analyze them, doing a density/ intensity plot over a sample lane. I am interested in the Intensity of the peaks (not necessarily the area, as there are often double peaks from not separated bands) and I want to know whether the two channels show peaks at the same position.
My beloved output would be: xy data of the 700and800 channel in one step
What I found out until now: plot profile of a composite allows me to save/export xy data, but in two steps and only from a line
The Gel analysis function in Image J makes horizontal means of an rectangle drawn around one sample lane, but cannot show me the two channels and I cannot export xy data.
Does anyone has any suggestions ?
Kind regards: anja

--
Anja Sieber
Medical Systems Biology
Charite - Universitätsmedizin Berlin
Institut für Pathologie
Chariteplatz 1, 10117 Berlin
Email: [hidden email]<mailto:[hidden email]>
Web: http://sys-bio.net/
Tel: [phone] (office)
Tel: [phone] (lab)


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[Tiago Ferreira-2]

On 2012.10.23, at 06:02 , Sieber, Anja wrote:
>
> Hi everyone,
> I have western blot pictures, either as a two channel picture or two separate pictures of the two channels (700/800nm). I want to analyze them, doing a density/ intensity plot over a sample lane. I am interested in the Intensity of the peaks (not necessarily the area, as there are often double peaks from not separated bands) and I want to know whether the two channels show peaks at the same position.
> My beloved output would be: xy data of the 700and800 channel in one step

The easiest way to obtain multiple line profiles on the same graph is to use the
Multi-Plot command.

1) Draw a line in channel 1 (or image 1), press "T" ("Edit>Selection>Add to Manager");
2) Move the c slider to channel 2 press "T" (or activate image 2 and press "Shift+E",
   "Edit>Selection>Restore selection")
3) In the ROI Manager, choose "Multi Plot" from the "more>>" dropdown menu.

HTH,
-tiago

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[Sieber, Anja]

Hi Tiago,
yes it helped. I had an older ImageJ version without the multi Plot function..
and what I have to say: one cannot make a multiplot of one roi in different pictures, but of different rois in one pic and then another plot with the same rois in another pic.
many thanks for this fast and helpful answer: anja

-----Ursprüngliche Nachricht-----
Von: ImageJ Interest Group [mailto:[hidden email]] Im Auftrag von Tiago Ferreira
Gesendet: 23. oktoober 2012. a. 17:20
An: [hidden email]
Betreff: Re: Licor Odyssey Western Blots

On 2012.10.23, at 06:02 , Sieber, Anja wrote:
>
> Hi everyone,
> I have western blot pictures, either as a two channel picture or two separate pictures of the two channels (700/800nm). I want to analyze them, doing a density/ intensity plot over a sample lane. I am interested in the Intensity of the peaks (not necessarily the area, as there are often double peaks from not separated bands) and I want to know whether the two channels show peaks at the same position.
> My beloved output would be: xy data of the 700and800 channel in one step

The easiest way to obtain multiple line profiles on the same graph is to use the
Multi-Plot command.

1) Draw a line in channel 1 (or image 1), press "T" ("Edit>Selection>Add to Manager");
2) Move the c slider to channel 2 press "T" (or activate image 2 and press "Shift+E",
   "Edit>Selection>Restore selection")
3) In the ROI Manager, choose "Multi Plot" from the "more>>" dropdown menu.

HTH,
-tiago

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ImageJ mailing list: http://imagej.nih.gov/ij/list.html

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[jharford]

"My beloved output would be: xy data of the 700and800 channel in one step"

Another possible solution for this would be to try Image Studio Lite software (www.licor.com/islite).  It is a free western blotting software and was developed by LI-COR.  You will be able to view both 700 and 800 channels overlaid, and report the density of each band.