tdTomato and LSM confocal

Posted by kspencer007 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/More-power-and-less-exposure-or-vice-versa-use-of-100-laser-tp3246685p3946951.html

Hello all;

>
> I have a colleague who is unhappy with the images of td-Tomato
> AI9 mice on our Fluoview 500. They first view their brain sections
> on our AX70, with mercury lamp illumination, a TRITC 535/50 EX filter,
> 610/75 EM filter, and like what they see. On our FV500 with 543 laser
> illumination (HeNe-Green 50% default AOTF increased to 90%), we have
> switched to a 560LP filter from the 560-600EM filter. This improved
> their results, but they still are disappointed in signal intensity. I
> recently had our system checked over by Olympus service, and we
> measured the laser output as 0.45mW out of the fiber (non-wavelength
> specific power meter), which is just off the best it's ever been. I
> have suggested opening the pinhole and sacrificing Z resolution for
> brightness.
>
> Would you have any other suggestions for increasing the intensity of
> their signal? I have explained filters and Z-thickness versus
> brightness. I don't know what to tell them, and they are faulting the
> system. From all spectra I can find, our system should be great as seeing this construct.

 Kathy Spencer