Is a CLSM absolutely required to take z stacks or can I try using a ordinary Compound Microscope

[dksamuel]

Dear all, the question may be so stupid and so I apologise at the beginning,
Is a CLSM (Confocal Laser Scanning Microscope) absolutely required to take z
stacks, or can I try using a ordinary Compound Microscope like Olympus BX41
etc, Can z planes be photographed using an ordinary consumer camera (Canon
A560)  or are motorized equipment required, can I try by by manual focusing
of some 10  - 20 slices and photograph each slice separately and use plugins
to assemble,  (Real reason is I want to play with stacks and learn volume
rendering by using volumeJ,  but I dont have a CLSM ready now or in the near
future)

Secondly has anybody tried ImageJ and Blender for rendering,

thanks Samuel, Virologist, Bangalore India

[Joel Sheffield]

The short answer is, "it depends".  That is, if the working distance
of your objective is short, you will be able to have a small depth in
focus at any time, and you could probably get away with a series of
steps.  Most microscopes have a calibration on the fine focus knob so
that you can obtains evenly spaced images.  You can also deconvolve
each of the images to remove out-of focus information, and then make
a stack of them.

You can see an example of a brightfield stack, not deconvolved, and
in color, on the left panel on our departmental web site:
http://www.temple.edu/biology.  .

The trick in making this image was to invert the original color
images, and use the stacks>3D routine set for Maximum brightness, and
then to invert the results again.

Date sent:       Wed, 12 Sep 2007 15:58:53 +0530
Send reply to:   ImageJ Interest Group <[hidden email]>
From:           Duleep Samuel <[hidden email]>
Subject:         Is a CLSM absolutely required to take z stacks or
can I try using a ordinary Compound Microscope
To:             [hidden email]

> Dear all, the question may be so stupid and so I apologise at the beginning,
> Is a CLSM (Confocal Laser Scanning Microscope) absolutely required to take z
> stacks, or can I try using a ordinary Compound Microscope like Olympus BX41
> etc, Can z planes be photographed using an ordinary consumer camera (Canon
> A560)  or are motorized equipment required, can I try by by manual focusing
> of some 10  - 20 slices and photograph each slice separately and use plugins
> to assemble,  (Real reason is I want to play with stacks and learn volume
> rendering by using volumeJ,  but I dont have a CLSM ready now or in the near
> future)
>
> Secondly has anybody tried ImageJ and Blender for rendering,
>
> thanks Samuel, Virologist, Bangalore India

--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
[hidden email]  
[phone], fax [phone]
http://astro.temple.edu/~jbs