Dear Matthias, dear all,
there is definitely no limitation to 10 steps in Lambda Scan (spectral scan) -
you can do as many steps as you like to do!
Please find attached a protocol how to do Lambda Scan with TCS NT software with
Leica TCS SP.
As Holly pointed out, the use of Lambda Scan in new software LCS is much more
user-friendly as it was in old software version.
Ulrike
(See attached file: LambdaScan.pdf)
_________________
Ulrike Tauer, Ph.D.
Application Specialist
Leica Microsystems Heidelberg GmbH
Am Friedensplatz 3
68165 Mannheim
Phone: +49 (0)621 7028 2704
Fax: +49 (0)621 7028 2780
mailto:[log in to unmask]
Stamatis Pagakis <[log in to unmask]> on 29.04.2001 19:15:06
Please respond to Confocal Microscopy List <[log in to unmask]>
To: [log in to unmask]
cc: (bcc: Confocal News/DEMAN/LMSCentral/Leica)
Subject: Re: Emission spectra with Leica TCS SP?
Hi
the way to do it is to set-up a number (I think 10 is the maximum the
software accepts) acquisition methods, each having identical parameters
except the emission bandwidth. Do then sequential acquisition with all
methods. You will collect a sequence of 10 images, each representing the
signal at the selected bandwidth. If you plot the integral within an ROI
for each of your images, you end up with a spectral plot of your
fluorophore. Hope it helps. Sorry for the brief description, but if you
need more info, please get back to me
regards
*-----------------*------------------*----------------------*---------------*
>Dr Stamatis Pagakis [log in to unmask] <
>Confocal and Image Analysis Laboratory Tel: +44 (0)20 8913 8675 <
>Membrane Biology Mobile: 07867 500029
>National Institute for Medical Research Switchboard: 020 89593666 x2621 <
>The Ridgeway Message: x2219, x2622 <
>Mill Hill, London NW7 1AA FAX: +44 (0)20 8906 4477 <
On Fri, 27 Apr 2001, Holly Aaron wrote:
> Hi, Matthias -
>
> It is possible. I think with the TCS software that you'd have to set up
> either a time series with different wavelengths, i.e., manually selecting
> different wavelength ranges, or some sort of sequential scanning, also
> setting your own. <i have never done it on the old software and my
> memory of that software is fading, but feel free to contact me offline and
> i can go into more detail on how to do that>
> The newer (much nicer) software from Leica - LCS - has a built-in function
> just for that - xy(z)-lamda (xyz-wavelength) - and it is quite elegant.
> You can specify the range, i.e. 500 to 600nm, the filter-width (10nm or
> whatever), and the number of steps to take (i.e. 10 to cover all
> wavelengths in that range). Then it's easy to plot out a graph from the
> data. I would highly recommend upgrading to the new software.
>
> Good luck,
> -Holly
>
>
> Matthias Heinemann wrote:
>
> > Hello!
> >
> > We wanted to determine the emission spectra of a dye with our
> > CLSM (Leica TCS SP with the software Leica TCS NT Version
> > 1.6.582).
> >
> > I am pretty sure that the our Leica representative had once
> > demonstrated a method (something like a time series with
> > changing detection wavelenghts ...). But, unfortunately I cannot
> > remember who is was done.
> >
> > I would be glad if someone could help me!
> > Thanks a lot in advance.
> >
> > Best Regards,
> > Matthias Heinemann
> >
> > --
> > -----------------------------------------------------------------------
> > Dipl.-Ing. Matthias Heinemann Aachen University of Technology (RWTH)
> > Department of Biochemical Engineering
> > Tel: +49 241 80-7072 Worringerweg 1
> > Fax: +49 241 8888-265 52074 Aachen, Germany
> >
> > [log in to unmask] http://www.biovt.rwth-aachen.de
> > -----------------------------------------------------------------------
>
> --
> Holly L. Aaron
> Berkeley Imaging Center
> http://imaging.berkeley.edu/
>
|
