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Re: not a confocal question - features of a widefield

Posted by Nuno Moreno on Dec 07, 2007; 6:18pm
URL: http://confocal-microscopy-list.275.s1.nabble.com/not-a-confocal-question-features-of-a-widefield-tp591195p591215.html

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This does not track cells that are moving in Z, right?

Engstrom, Lars wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> Nikon has a Perfect Focus System I believe for their TE2000 series. It
> uses an LED (770nm) to focus on the coverslip and adjust Z prior to
> image acquisition.
> http://nikonusa.com/fileuploads/pdfs/Instruments/TE2000_PFS.pdf
>  
> I haven't tried it but sounds similar to what you are looking for and
> could use their system as an example for building your own.
>  
> -Lars
>
> ------------------------------------------------------------------------
> *From:* Confocal Microscopy List [mailto:[hidden email]]
> *On Behalf Of *Zoltan Cseresnyes
> *Sent:* Friday, December 07, 2007 9:50 AM
> *To:* [hidden email]
> *Subject:* Re: not a confocal question - features of a widefield
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> I've been thinking about building a system like that ever since one of
> our users started doing overnight recordings and his samples sometimes
> drift vertically and thus would need an autofocus system to correct for
> that.  If you could tap into the Z-controller circuit of your system,
> you would be able to drive that with a signal that's proportional to the
> vertical drift.  The drift could be scaled with e.g. calculating average
> intensity during a Z-stack from a certain area of the image that's
> fairly thin.  Similar logic would work for the autoexposure as well.  
> Want to try it?
>  
> Zoltan
>
> On Dec 7, 2007 4:41 PM, Nuno Moreno <[hidden email]
> <mailto:[hidden email]>> wrote:
>
>     Search the CONFOCAL archive at
>     http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>     Autoexpose will bleach everything, right?
>
>     Regarding the adaptative focus that I mentioned before, there are
>     commercial system that with minimum light and before an acquisition
>     "measure" the cell position and adapt the focus. But this is an half
>     adaptation. It could be that it does not need to readjust the focus.
>
>     What I was counting with would be after the acquisition, if it is out of
>     focus, it make the adjustment base in some kind of sensitivity
>     parameter. This could be after 10 time points but it might be that it
>     would never need such adjustment.
>
>
>     About the intensity variations I'm not talking about post processing
>     adjustments. If it gets saturated there are no post processing that can
>     help you.
>
>     Regards,
>     NM
>
>
>
>
>
>     Shalin Mehta wrote:
>      > Search the CONFOCAL archive at
>      > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>     <http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>Dear Nuno,
>      >
>      >  Wouldn't auto-exposure on cameras suffice for maintaining constant
>      > intensity?
>      >
>      > Apparently most of the commercial adaptive optics systems are geared
>      > towards astronomy. Perhaps you have known this already:
>      > http://cfao.ucolick.org/
>      > Interesting to note that James Webb space telescope will have
>     hardware
>      > and intelligence for adaptive optics evolved from algorithms
>     developed
>      > for correcting aberrations for hubble telescope.
>      >
>      > Regards,
>      > Shalin
>      >
>      >
>      > On Dec 7, 2007 10:43 PM, Nuno Moreno <[hidden email]
>     <mailto:[hidden email]>
>      > <mailto:[hidden email]
>     <mailto:[hidden email]>>> wrote:
>      >
>      >     Search the CONFOCAL archive at
>      >     http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>      >
>      >     Does anyone knows any commercial widefield SYSTEM that makes an
>      >     adaptative focus. And I mean adaptative (follows the cell.
>      >
>      >     The other feature is a commercial system that keeps
>     intensities, i.e.,
>      >     if you have something with different protein expression
>     levels over
>      >     time, the system will correct the exposure time so that at
>     the end the
>      >     intensities are constant.
>      >
>      >     Many thanks,
>      >     --
>      >     Nuno Moreno
>      >     Cell Imaging Unit
>      >     Instituto Gulbenkian de Ciência
>      >     http://uic.igc.gulbekian.pt <http://uic.igc.gulbekian.pt/>
>     <http://uic.igc.gulbekian.pt <http://uic.igc.gulbekian.pt/>>
>      >     http://www.igc.gulbekian.pt <http://www.igc.gulbekian.pt/>
>      >     phone +351 214464606
>      >     fax   +351 214407970
>      >
>      >
>      >
>      >
>      > --
>      > ~~~~~~~~~~~~~~~~~~~~~~~~~
>      > Shalin Mehta
>      > Graduate Student in Bioengineering, NUS
>      > mobile: +65-90694182
>      > blog: shalin.wordpress.com
>     <http://shalin.wordpress.com/><http://shalin.wordpress.com
>     <http://shalin.wordpress.com/>>
>      > ~~~~~~~~~~~~~~~~~~~~~~~~~~
>
>     --
>     Nuno Moreno
>     Cell Imaging Unit
>     Instituto Gulbenkian de Ciência
>     http://uic.igc.gulbekian.pt <http://uic.igc.gulbekian.pt/>
>     http://www.igc.gulbekian.pt <http://www.igc.gulbekian.pt/>
>     phone +351 214464606
>     fax   +351 214407970
>
>
>
>
> --
> --
> Zoltan Cseresnyes
> Facility manager, Imaging Suite
> Dept. of Zoology University of Cambridge
> Downing Street, Cambridge
> CB2 3EJ    UK
>
> Tel.: (++44) (0)1223 769282
> Fax.: (++44) (0)1223 336676

--
Nuno Moreno
Cell Imaging Unit
Instituto Gulbenkian de Ciência
http://uic.igc.gulbekian.pt
http://www.igc.gulbekian.pt
phone +351 214464606
fax   +351 214407970