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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