Posted by
Steffen Dietzel on
URL: http://confocal-microscopy-list.275.s1.nabble.com/white-light-laser-tp7590046p7590066.html
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Dear Konstantin,
apart from things that were already mentioned by others, the fact that
they are pulsed is an advantage and disadvantage. Gated detection with
the HyDs is an extremely cool feature to get rid of reflected background
noise. Plant people also seem to love this since you can gate out the
very short lifetime chlorophyll fluorescence. On the downside, you can
excite any fluorochrome only every 12.5 seconds, so you may run into
saturation faster than with a cw laser. Another downside is that the
excitation line is not as narrow as with single lasers. In other words,
it seems that the AOTF is not 100% tight against neighboring lines. In
spectral detection we advise our users to keep 10 nm between excitation
and detection. This is a very cautious approach, and if you are careful
and know what you are doing you can get closer. But as a general rule
we'd prefer to be on the safe side for the HyDs. Now you could argue
that the gated detection reflection suppression is a problem that you
wouldn't have without the WLL, but I don't think so. Some samples
somehow create a strong reflection near the coverslip that I don't think
is coming from the WLL, I know this pattern also from single line systems.
Also, the option of exciting with 670 opens up another channel above 630
excitation that you don't reach with a 488/552/594/633 battery of lasers.
Maybe in some experiments it is helpful to know that there will be zero
misalignment between the laser lines (except for the 405 and the WLL)
Just my two euro-cents.
Steffen
Am 21.10.2019 um 15:28 schrieb Konstantín Levitskiy:
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> Dear Microscopists,
>
> Do you know if any other company, besides Leica, include white light laser
> in their confocals. Or if any other technology can replicate the free
> selection wavelength laser light? On the other hand, what are disadvantages
> of the WLL?
>
> Best regards,
>
> Dr. Konstantín Levitskiy
>
> Servicio de Microscopía
>
> InstitutodeBiomedicinadeSevilla - IBiS
>
> Email: <mailto:
[hidden email]>
[hidden email]
>
> Web: <
http://www.ibis-sevilla.es/> www.ibis-sevilla.es
>
>
>
--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging
Großhaderner Straße 9
D-82152 Planegg-Martinsried
Germany
http://www.bioimaging.bmc.med.uni-muenchen.de