Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Glicerin-labeling-tp7265984p7269081.html
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Hi Arvydas,
Highly reminiscent of the LSM510 I manage(d) here in Miami (PC died
recently - finally able to decommission it - if you or anyone else needs
parts, make UM an offer ... I am putting on a digital camera system on
the Axiovert 200M stand, though only may be able to use the right side
camera port - left side may have some factory installed features
specific for the LSM light path).
It took Zeiss a lot longer than I would have liked, but one of their
field service engineer's greatly improved out LSM510 by removing the ~25
mm of dust that had assumulated in the dust trap under the electronics box.
See this paper for the nice, the bad and the ugly of other confocal's:
Quality assurance testing for modern optical imaging
systems. </pubmed/21477410> Stack RF, Bayles CJ, Girard AM, Martin K,
Opansky C, Schulz K, *Cole RW*. Microsc Microanal. 2011
Aug;17(4):598-606. Epub 2011 Apr 11. PMID: 21477410
In other news ... our Leica SP5 inverted microscope had a very dim PMT3.
This week, Leica field service (thanks Jim C and Gene B) replaced the
PMT - it is now the brightest. They will be visiting in a few weeks to
replace three other PMTs in that SP5, plus the 3 standard PMTs in our
MP/SP5/FCS/FLIM - having a service contract is a good thing. Each
machine is close to 5 years old and have been sustaining close to 1,000
hours of use (Sp5 inverted use now up thanks to retiring the old LSM510).
George
On 2/8/2012 12:34 PM, Arvydas Matiukas wrote:
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> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> Dear list,
>
> Please advice what is the most efficient/cheap way to deal
> with confocal laser instability. Our LSM 510 is 7 years old
> but Ar and Hene 543 lasers have been replaced two years
> ago and have been used no more that 2000-2500 hrs.
> Users started complaining about 488nm and 543nm laser
> line stability, and I measured it using Chroma fluorescent
> plastic slides and 10x objective. The frame- averaged fluorescent
> signal is reduced by 40-50% in 1hr after laser start. The decline
> curve is quite steady after first 3-5 min with some fluctuations.
>
> The most disturbing are intermittent large scale signal fluctuations (2-4 fold)
> occurring on about a minute timescale. They kill dynamic (e.g. FRET) or
> long-term imaging. They happen irregularly, on average once in 4-5 days,
> and always after 2-3 hrs after the lasers are started.
>
> In a related post several years ago laser cooling, laser polarization, AOTF cooling,
> and AOTF driver stability were listed as the key factors. I am quite confident that cooling
> is not an issue as the fans within the equipment boxes are working well, and whole room
> has a powerful local conditioning system which maintains ambient temperature very stable.
> Zeiss engineer checked the fibers recently, and suggested that maybe AOTF drivers are
> not stable. As we are not on service contract and have to pay for every replacement I
> would be interested to do more testing myself to identify the failing component.
>
> Any suggestions/advices are very welcome. I can provide the measurement curves if
> somebody is intersted (contact me off list).
>
> Thanks,
> Arvydas
>
>
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: 315-464-7997
> fax: 315-464-8014
> email:
[hidden email]
>
>
--
George McNamara, PhD
Analytical Imaging Core Facility
University of Miami