Posted by
Arvydas Matiukas on
URL: http://confocal-microscopy-list.275.s1.nabble.com/xyzt-image-tp7580102p7580120.html
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Hi Pascal,
Thanks for the suggestion to acquire both DAPI and FITC signals
in 2P mode. Actually I typically use double DAPI/FITC
configuration with FITC channel disabled.
I agree that by exciting both DAPI and FITC at the same 2P
wavelength provides a perfect colocalization. I will tweak
the settings to get less noisy images.
I did not initially use FITC acquisition in 2P mode because this mode
yields more noisy signal compared with confocal. Additionally,
LSM software was giving incorrect optical slice for 2P mode
which confused me.
Best regards,
Arvydas
>>> Pascal Weber <
[hidden email]> 4/4/2013 2:10 AM >>>
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In fact for colocalization signals, DAPI and FITC, just use the laser 2P. You'll
always have a perfect colocalization. The fluorescence comes as a single dot.
Why would you want to use two different lasers?
For the LSM510 the only way to adjust the filling of the rear lens you have to
ajust the beam size with a lens. I did it and i can modulate the beam size and the
uniformity of the field.
Regards, Pascal