Re: Z sections

Posted by Christian Liebig on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Z-sections-tp1142113p1300623.html

Hi Carol,

what the Leica software does (AFAIK):
- you choose the number of layers you want (dropdown menu or over the
"others" dialogue, doesn't matter)
- the software divides the stacksize chosen by you with this number and
gets a value for your stepsize (if you choose a stepsize from the
"others" dialogue start from here)
- the software adapts the stepsize value to the nearest stepsize the
z-motor can handle
- then the software multiplies the new stepsize with the number of
layers and there you have the new z range for your stack.

This is just 3rd party information, maybe it would be better if someone
from Leica would comment on this.

Kind regards,

Christian



Carol Norris wrote:

> Hi Matt,
>
> Just some advice for after you've decided how many sections to use.
> Don't try to use one of the other preset #sections that appear when
> you click on the "sect" button.  I found that this causes the
> coordinates for the top and/or bottom of the stack to change. I tested
> this by imaging the same stack three times in a row at different
> preset section numbers, and found that the top of the stack had
> changed by 3 microns.  Leica had no explanation for this, they just
> said to select "others,"  which brings up a window in which you can
> enter either a section spacing or the number of sections.  
>
> Regards,
> Carol
>
> On Oct 3, 2008, at 10:40 AM, Matthew Pearson wrote:
>
>>
>> Hi all,
>>
>> I have a question concerning Z sectioning.  We have a Leica SP2 and
>> when set the top and bottom range for a Z stack, the software
>> automatically calculates the optimised number of sections, which is
>> nearly always over 100!  This is however not practical as the stacks
>> take too long to acquire.  Can anyone shed some light on how the
>> software decides how many slices are necessary and if I haven't got
>> time to take 100, how can I work out what will be a sufficient number.
>>
>> Thanks for your help in advance.
>>
>> Matt Pearson.
>>
>>
>
>
>
> Carol E. Norris, Ph.D
>
> Facility Scientist
>
> Flow Cytometry/Confocal Microscopy Facility
>
> Biotechnology/Bioservices Center
>
> University of Connecticut Unit 3149
>
> 91 N. Eagleville Rd
>
> Storrs, CT 06269-3149
>
>
> Phone [phone]
>
> Fax [phone]
>
>

--



Christian Liebig, PhD

FILM - Facility for Imaging by Light Microscopy
- Facility Assistant -
Sir Alexander Fleming Building, desk 403
Imperial College London / South Kensington
Exhibition Road
London SW7 2AZ
UK

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