>Hello all,
>
>We have a Zeiss LSM410 confocal laser scanning microscope, but we (the
>laboratory confocal users) are having trouble understanding how to use
>some of the software applications.  For example, I wanted to get a simple
>picture with most of the image (a chick embryo somite) in gray scale,
>except for the fluorescent labelled area (myotome cells) which would be
>colored (red or green).  However, after spending hours, without success,
>trying to figure out how to do this, I finally moved the files to Adobe
>Photoshop to get the picture.  I am frequently running into these kinds of
>blocks because I find the LSM410 user manual difficult to understand for
>using the software to analyse images.  Does anyone have, or know of, a
>user manual for the Zeiss LSM410 that clearly explains how to carry out
>image analysis using the microscope software?
>
>Thank you,
>
>Wilfred F. Denetclaw Jr., Ph.D.
>Dept. of Anatomy
>UC San Francisco
>San Francisco, CA  94143-0452
>(415)476-1530
 
Unfortunately Zeiss's manual is not very simple to follow.  But for your
paricular problem, if I understand you correctly,  it sounds like that you
need to put a phase contrast or DIC image in  say the blue chanel.  Then
put your labeled image in the red or the green (depending on the label you
are using: FITC, Rhodamine, TexasRed, etc.) chanel.  After this, under the
image menue go to "Arithmatic Operations" submenue and add the gray image
(phase or DIC) to each of the chanels.  Then when you combine the images
under RGB,  you should see the gray phase background with the fluorescent
label superimposed on top of it.  If done right, the image is fantastic.
You can call Brad Whitman at Zeiss (304-599-8018) and he can walk you
through the procedure. Or you can call me if you like and I will give you
complete instructions or even walk you through it.
 
Good luck.
 
Sincerely,
 
Kamiar Moin, Ph.D.
Assistant Professor of Pharmacology
Wayne State University School of Meddicine
Detroit, MI 48201
 
Tel:  (313)577-0514
        (313)577-1112
FAX: (313)577-6739
E-mail:  [log in to unmask]