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Hi everybody,

Just two questions about developement of a quite old Leica TCS-NT
Confocal, the one that uses conventionnal filters in the detection part
of the scanhead.

1) We would like to image eCFP and eYFP, not necessarily
simultaneously. We have an Ar/Kr and we currently have the 458, 488,
568 and 647 nm lines working (our AOTF can support up to 4 lines). For
eYFP eGFP settings are quite OK (488 excitation, RSP500 dichroic, BP
525/50 emission). I tried to image eCFP using 458 nm excitation with
the same dichroic and emission filters but I got nothing. I was
wondering if I could get a dichroic with <500nm in order to detect eCFP
fluorescence at lower wavelength. For simultaneous detection, is there
any DD458/488 ? or should we get the 514nm laser line working for eYFP
and use a DD458/514 ? I tried to have a look at the litterature but
authors rarely go in such optical details in the material & methods...

2) We just got a nice XY automated stage, but the only Leica LCS macro
I got can memorize XY position but not the motorized Z position. Do
anyone have a macro that memorizes XYZ position for multiple time-lapse
experiments ? More general, do anyone know where I can find a reference
for all objects used by the Leica TCS VBA macro programming ?

Thanks,


-- Christophe LETERRIER -------------------

Ph D Student
Neuronal Receptors Team
Neurobiology Laboratory
ESPCI-CNRS UMR 7637
10, rue Vauquelin 75231 Paris Cedex 05

Tel 01 40 79 51 84
Fax 01 40 79 47 67
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