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May 2008

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Subject:
Re: Correcting misfocusing deviation in confocal FRET imaging.
From:
Michael Weber <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 2 May 2008 14:10:02 +0200
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

David,

you can get 0.17 µm multicolor beads from Invitrogen. There size is 
below the common resolution limit of light microscopy, so you can use 
them to test and calibrate your setup. Get them in solution and make 
your own slides, the prepared ones are quite expensive.

Michael


david anchel wrote:
> Search the CONFOCAL archive at 
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi there,
> 
> I too have the same problem with aligning the Z focus of my NIR and VIS 
> lasers.  Where can we get these .1um multicolour beads?
> 
> Dave Anchel,
> Toronto
> 
> <html><DIV>
> <DIV></DIV></DIV></html>
> 
>  > Date: Thu, 1 May 2008 17:27:43 -0400
>  > From: [log in to unmask]
>  > Subject: Re: Correcting misfocusing deviation in confocal FRET imaging.
>  > To: [log in to unmask]
>  >
>  > Search the CONFOCAL archive at
>  > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>  >
>  > Suggest testing the system with fine Z series of 0.1 um multicolor beads.
>  >
>  > >
>  > > In a paper, Biophysical Journal, vol 86, p2517, 2004, the work 
> mentioned
>  > > several factors needed to be corrected when perform FRET in laser 
> scanning
>  > > confocal, by sensitized emission. One of them is to correct misfocusing
>  > > deviation (page 2521). This error comes from two main sources. One is
>  > > chromatic aberration, which we are quite familiar. The other is the 
> slight
>  > > differences in the collimation of the laser beams.
>  > >
>  > >
>  > >
>  > > Any comments?
>  > >
>  > > Are there ways to test the performance of the system?
>  > >
>  > > Thanks
>  > >
>  > >
>  > >
>  > > Hao-Yang Liu
>  > >
>  >
>  >
>  > _________________________________________
>  > Michael Cammer http://www.aecom.yu.edu/aif/
> 
> ------------------------------------------------------------------------
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