LU,Pin-
 
You don't say what the source images are or what version of Photoshop you
are using. For full frame BioRad MRC600 files and Photoshop 3.o, the
instructions follow. For other options, contact me and I'll be glad to
help.
 
First import the confocal images into Photoshop using "Open..Select all
files..file type RAW"
 
The "Raw Open" screen prompts you for Width (768) Height (512) Channels
(1) and Header (76). Then say OK and OK again when told that the
specifications are smaller than the actual file size (It is smaller due
to footer info at the end of the file).
 
Do the same for whichever files you are trying to merge. A hint is that
you shoud open them in Red-Green-Blue order since it will save a step
later. If you are not merging 3 channels, then you will need to duplicate
one of the channels (Image..Duplicate) and then fill the duplicated image
with black (0). At this point you should have three grey-scale images.
There are a couple of ways to go from here. My favorite is to convert one
of the grey-scale images to RGB using the Mode...RGB command, lets
say the "red" image. Then go to whichever image is to be "green" and
select all (Command-A on a Mac or Cntrl-A on a PC). Do a copy (command-
or cntrl-C). Go back to the Red window and select the green channel
(Command-2 on a Mac) and paste the green image in (Command-V). Then go to
the blue channel, select all, copy and go to the red window, select the
blue channel (command-3) and paste. Command-0 (that is a zero) will show
you the merged image.
 
If you are going to make color prints, you may wish to "enhance" the blue
since pure blue does not print well on most printers. To do this, use the
calculate command (Image...Calculations) to add 50% of the blu into the
green channel which will make the blu much lighter (close to Cyan which
will print much better). In the Calculations menu, choose the blue
channel as source 1, the green channel as source 2, for Blending choose
add, Opacity 50% and Result the Green channel. I normally save this as a
seperate file from my true 24-bit color merge. The original 24-bit file
should be OK for slides, but we find the Enhanced version far superior
for printing.
 
I hope this is clear. Let me know if you come up with something easier.
In Photoshop Version 3.0 or higher you can create short-cuts using the
commands window for many of theses tasks.
 
Regards-
 
________________________________________________________________________________
 
 
Paul Goodwin
Image Analysis Lab
FHCRC, Seattle, WA
 
On Fri, 27 Oct 1995, LU Pin wrote:
 
> Hello, everyone,
>
> I remember several monthes ago someone posted a detailed step-by-step
> tutorial of how to merge image files in Photoshop. I have searched
> confocal mailing list archive without success. Will someone please re-post
> it and mail it to me? Thank you.
>
> Lu Pin
> Oct-27, 95
>
> ====================================================================
> LU, Pin                       +  Phone:  (852)-2358-7312
> Department of Biology         +  Fax:    (852)-2358-1559
> The Hong Kong University of   +  e-mail: [log in to unmask]
>   Science & Technology        +          [log in to unmask]
> Kowloon, Hong Kong            +  WWW:    http://www.ust.hk/~bolupin
> ====================================================================
>