A standard lamp calibration is necessary if you'd like to record a meaningful spectrum of your dye. It allows you to correct for the wavelength-dependent transmission of your system. As discussed many times in this list, all of the optical components in the system absorb light at some wavelengths. A standard lamp allows you to correct for these absorptions (instrument response curve). Also, if one of your optics begins to fail it should show up in the instrument response curve.
Sarah
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Sarah Locknar, Ph.D.
Director, Neuroscience COBRE Imaging / Physiology Core
College of Medicine, University of Vermont
E015 Given Building
89 Beaumont Ave.
Burlington, VT 05405
802-656-0413
802-656-8704 (fax)
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Guy Cox
Sent: Monday, August 09, 2004 7:47 PM
To: [log in to unmask]
Subject: Re: Spectroscopy Confocal QA
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Sounds interesting - please do send me a copy.
But my question is why is this necessary? You have absolute standards built in the in the form of your CW lasers (6 lines on our system) - their position shows how accurately your system is calibrated and the FWHM shows the resolution. What more do you need?
Guy
Quoting Robert Zucker <[log in to unmask]>:
>
> At the recent microscopy meetings (MM2004) in Savannah a paper by R
> Zucker and J. Lerner was presented entitled “Confocal Microscopy
> System
> Performance: Foundations for Measurements, Quantitation and Spectroscopy
> â€
>
> This Confocal QA spectroscopy test was developed to measure spectral
> performance and to serve as guidelines for investigators to assess
> both the performance of their instruments as well as the quality of
> their data. This spectral characterization test is well suited to all
> wavelength dispersive CLSM systems including the Leica SP series (SP),
> the Zeiss LSM510 Meta (Meta) and Olympus FV1000 confocal microscopes.
> We used an inexpensive, eye-safe, battery operated, multi-ion
> discharge lamp (MIDL) (LightForm, Inc., Hillsborough NJ) containing
> mercury ions and inorganic fluorophores as an absolute reference light
> source because it emits stable, reproducible, peaks between 400 and
> 650 nm. It is worth applying this test to your systems.
>
> An abstract describing this presentation has been published and is
> available on request.
>
> Bob
>
>
> Robert M. Zucker, PhD
> U.S. Environmental Protection Agency
> Office of Research and Development
> National Health and Environmental Effects Research Laboratory
> Reproductive Toxicology Division, MD 72 Research Triangle Park, North
> Carolina, 27711
> Tel: 919-541-1585; fax 919-541-4017
> e-mail: [log in to unmask]
--
Associate Professor Guy Cox
Electron Microscope Unit, F09
University of Sydney NSW 2006
+61 2 9351 3176
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