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Hi Pedro,
On our Biorad 1024UV system, using the Nikon 60x NA1.4 PlanApo objective, the final pixel size is 310nm x 310nm. Assuming optimal pixel size of 80nm for Nyquist sampling to resolve 200nm in x,y then the maximum "useful" zoom is 310/80 = 3.8. On different confocal systems the final pixel size could be quite different. I have always understood this is how Biorad calculated their optimal zoom value for that objective.
Cheers
Paul
Dr Paul Rigby
Senior Lecturer
Centre for Microscopy, Characterisation and Analysis (M510)
The University of Western Australia
35 Stirling Highway
Crawley WA 6009
________________________________
From: Confocal Microscopy List on behalf of Pedro J Camello
Sent: Sun 10/02/2008 8:30 AM
To: [log in to unmask]
Subject: Re: Best parameters for optical slicing
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Yes, nm.
Julio, you are right about lateral resolution. My reasoning is that
although the optical performance does not depend on the structure, 150 nm
is supposed to be good enough to sample structures of 500 nm. I would like
to avoid too much oversampling.
I donīt know how Biorad calculated that 2.6 zoom is the maximum advisable
for this objective (it is a shame, but I never worried before about
sampling rate, resolution, etc)
Many thanks for your help.
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello All,
>
> To avoid more confusion, you both meant nanometers, not micrometers, when
> talking about xy-, z-resolution and z-stepsize, correct?
> E.g., "optimal" xy resolution is 200 nm, not microns, with pixel size at
> 80
> nm with Nyquist, 160 nm z-stepsize, etc.
>
> Zoltan
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