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Dear Holly,

If I understand correctly, you already have a RGB overlay (the RGB image)
and you want to create a combined overlay with the 8bit grayscale. In this
case you first have to separate the RGB into the 3 color channels of the RGB
(in a RGB file a red (R), green (G) and blue (B) - usually they have 8bit
each so you end up with a 24bit RGB file). Afterwards you can combine the 4
8bit images to a RGB+gray overlay using the "RGB gray merge" plugin
(http://rsb.info.nih.gov/ij/plugins/rgb-gray-merge.html), which is also
included in the WCIF ImageJ bundle by Tony Collins
(http://www.uhnresearch.ca/facilities/wcif/download.php) in the group "Color
Functions".

In short:
- open the brightfield file and the RGB file
- separate the RGB into the 3 channels with image-> color -> RGB Split
- use the plugin "RGB gray merge" to combine all 4 images into one overlay. 

Best wishes,

Stefan



 

-------------------------
Dr. Stefan Terjung
EMBL, Meyerhofstr.1
D - 69117 Heidelberg
Cell Biology/Biophysics Program
Advanced Light Microscopy Facility
Phone ++49-6221-3878-467 FAX-242
www.embl.de/almf/
www.embl.de/eamnet/ 



> -----Original Message-----
> From: Confocal Microscopy List 
> [mailto:[log in to unmask]] On Behalf Of holly snyder
> Sent: Dienstag, 17. Januar 2006 04:08
> To: [log in to unmask]
> Subject: overlay in Image J
> 
> 
> Search the CONFOCAL archive at 
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> 
> Hi,
> 
> I have been using overlay in image J to merge confocal
> images and it works great.  However, I can't seem to
> overlay an image taken with brightfield and
> fluorescence.  One image is a 8 bit greyscale image
> and the other is a 32 bit RGB.  I keep getting the
> error mismatch type and I don't know what that means. 
> Is it because one is an 8 bit image and the other is a
> 32 bit image?  Thanks if anyone can give me any input.
> 
> Holly
> 
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