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Has your META been calibrated ?


On Nov 8, 2007, at 7:40 PM, Oky wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear all,
>
> * What is the difference of the detector of CLSM (Zeiss LSM 510 META)
> and fluorescence spectrometer (microplate reader, Molecular Devices
> spectraMax)? Or excitation source?
> For the microplate reader, I used the excitation wavelength at 350 nm
> and collected fluorescence intensity between 360 - 720 nm. The
> emission wavelength interval was 2 nm.
> For the META detector, UV laser at 364 nm was used for excitation and
> used all 32 channels for collection of emission. The emission
> wavelength interval was given as 10.7 nm.
>
> I have scanned some wood cell wall utilizing both instrument. I
> expected very similar spectra from both. The microplate reader seems
> to generate similar spectra to those in literatures, but the spectra
> from the META detector were very different from those of the
> microplate reader. The spectrum from the META detector consists of
> four major peaks with successively decreased intensity. If I connect
> only the peaks, it would be similar to the spectrum from the
> microplate reader.
>
> I have been trying to figure out what causes the difference. I want to
> make sure whether the characteristics of the instrument or something I
> have done incorrectly cause the differences in the spectra from both
> instrument. Here are a few possible reasons I could draw.
> 1. differences in light source and optical setup in each instrument
> 2. beam splitter (UV/488/543/630; the number may not be exact, but
> close) in Zeiss LSM 510 META
> 3. reflection between cover glass and sample surface
>
> Thanks.
>
> -- 
> Ohkyung Kwon, Ph D
>
> http://www.meso.or.kr/portfolio
> http://www.wpskorea.org