Posted by
Vitaly Boyko on
URL: http://confocal-microscopy-list.275.s1.nabble.com/7th-Course-on-Optical-Microscopy-Imaging-for-Biosciences-20-24-April-2015-IBMC-Porto-tp7583912p7583955.html
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Hi Craig,
thank you for your reply. How would one image Alexa 790/Cy7 together with let say CeFP/dapi, GFP through the one photon confocal system (with Perkin Elmer APDs as detectors). Are there many IR APOCHROMAT lens that are corrected between 400 nm (tough, I guess, at least 500 nm) and 900nm at 40x NA 1.0-1.1 or 60x NA 1.2?Multi-photon collect scattered (non-linear, non-chroma corrected) light via NDDs...
Vitaly
On Monday, June 29, 2015 6:16 PM, Craig Brideau <
[hidden email]> wrote:
Your question is fairly vague, but here are some general answers: With 2 photon the important thing is how well the laser is focused. This means your objective has to be well corrected for the NIR beam from the laser. The resulting visible fluorescence is directed towards wide-aperture PMTs, which don't care about aberration very much.For single-photon confocal, you have to get your fluorescence through the confocal pinhole, which will be *very* picky about aberration. Since you are trying to get the light through a very small aperture any aberration will cause signal loss.
Craig
On Mon, Jun 29, 2015 at 4:04 PM, Vitaly Boyko <
[hidden email]> wrote:
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Dear List,
I would greatly appreciate if someone could shed light on the extent of lateral and axial chromatic aberrations correction, and its limitations in the VIS portion of spectrum when image has been projected into the non-descanned detectors (one and two photon imaging)?
Many thanks in advance.
Vitaly