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Hello Graham,

On the 510 you can do the following:
-create 2 tracks (or configurations, depending if you wish to change any
parameter but the colors), (a) for the 'blue & red', (b) for the
'green'. 
-Mark stack top & bottom in the z settings dialog.
-press the 'median' button from the z settings dialog and acquire a
single plane with condition (a) then
-acquire a stack selecting condition (b).

You could then project the (b) or a subset of it and merge it with the
(a), though this will produce a complicated picture, which needs quite
some explanation.

You can automate the procedure using the 'multi-time' macro downloadable
from the Zeiss microscopy website, but I would not do it for just one
set of data, because the macro is quite difficult to handle.

The above mentioned procedure does only make sense if the green channel
is bleaching too quickly than to allow for taking a stack as well in
this channel.

Hope that helps, jens.


---
dr. jens rietdorf 
head microscopy
novartis research foundation
friedrich-miescher-institute, wro1066.2.16 
maulbeerstr.66, 4058 basel, switzerland
couriel:rietdorf(at)fmi(dot)ch
fon: +41616975172
fax: +41616973976



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Dr. Graham Wright
Sent: Thursday, September 11, 2008 3:31 AM
To: [log in to unmask]
Subject: Different Z setting per channel - possible on Zeiss/Leica?

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Hello,

Here's question that I've been asked that doesn't seem possible to do,
but I 
wonder if anyone has already done it? One of our users has a fixed
sample 
with 3 fluorophores (for ease of explanation blue, green and red) and
would 
like to image all three channels, but with different z setting depending
on 
the channel. She would like a single medial optical section of the blue
and 
red channels, but a full z-series from the green channel.

We have both Zeiss (from the LSM 510 series) and Leica (SPE and SP5) 
confocals available to us.

I know this is easy by doing 2 different scans: 1 2-channel scan of the
blue 
and red, then a second scan of a z-series of the green, BUT is it
possible 
to do this at the single click of a button to save time and user input.
It 
is possible on the SP5 using the live data mode, but this scope is
really 
reserved and heavily used for live cell imaging. Our SPE doesn't have
live 
data mode and therefore applies any Z settings to all sequential scans 
(generating unnecessary quantities of data and taking too long).

Any suggestions welcome,
Thanks,
Graham