overlaying images

[Ramon Cajal]

Does anybody knows who to overlay 2 different images? I have 2 images
of the same object stained with two different fluorochromes and I
need to overlay them

Thanks a lot
Ramon

[John Oreopoulos]

http://rsbweb.nih.gov/ij/docs/menus/image.html#color

RGB Merge:
Merges 1-4 greyscale images or stacks into an RGB image or stack.  
Select *None* to keep a channel empty (filled with 0). Check "Create  
Composite" to convert 2-4 grayscale images or stacks into a composite  
image or hyperstack. Check "Keep Source Images" if you wish to keep  
the originals.


On 28-Jan-10, at 10:12 PM, Ramon Cajal wrote:

> Does anybody knows who to overlay 2 different images? I have 2  
> images of the same object stained with two different fluorochromes  
> and I need to overlay them
>
> Thanks a lot
> Ramon

[Marko Usaj]

I have another questions about image overlaying.
I wondering how you solve the issues of uneven staining of object of interest. The problem occurs, when you try to detect by visual inspection an overlaying colors (on the same object) but one color has much higher gray scale values than other one what has for consequence wrong interpretation.

Ok, one option is to threshold images, but I don't want to lose all dynamic range of fluorescent images. Other options is histogram equalization (BTW which function is preferred for this task in imageJ?).
 
Is here any other (more nobel) solution and you are willing to share with me (us)?

Best regards,
Marko




-----Original Message-----
From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of John Oreopoulos
Sent: Friday, January 29, 2010 4:52 AM
To: [hidden email]
Subject: Re: overlaying images

http://rsbweb.nih.gov/ij/docs/menus/image.html#color

RGB Merge:
Merges 1-4 greyscale images or stacks into an RGB image or stack.  
Select *None* to keep a channel empty (filled with 0). Check "Create  
Composite" to convert 2-4 grayscale images or stacks into a composite  
image or hyperstack. Check "Keep Source Images" if you wish to keep  
the originals.


On 28-Jan-10, at 10:12 PM, Ramon Cajal wrote:

> Does anybody knows who to overlay 2 different images? I have 2  
> images of the same object stained with two different fluorochromes  
> and I need to overlay them
>
> Thanks a lot
> Ramon

[Janne Hyötylä]

Hi,

You could try using quantitative colocalization analysis:

http://www.macbiophotonics.ca/imagej/colour_analysis.htm
http://pacific.mpi-cbg.de/wiki/index.php/ColocalizationAnalysis

Cheers,
Janne

On Fri, 29 Jan 2010 10:01:49 +0100, Ušaj, Marko <[hidden email]>  
wrote:

> I have another questions about image overlaying.
> I wondering how you solve the issues of uneven staining of object of  
> interest. The problem occurs, when you try to detect by visual  
> inspection an overlaying colors (on the same object) but one color has  
> much higher gray scale values than other one what has for consequence  
> wrong interpretation.
>
> Ok, one option is to threshold images, but I don't want to lose all  
> dynamic range of fluorescent images. Other options is histogram  
> equalization (BTW which function is preferred for this task in imageJ?).
> Is here any other (more nobel) solution and you are willing to share  
> with me (us)?
>
> Best regards,
> Marko