Which pinhole size to use in the Leica SP2 confocal microscope?
I am using Leica TCS SP2 confocal microscopy. The pinhole setting is 1 Airy, which is about 82um. These information are obtained directly from Leica software. But When I was using Huygens essential, it told me the pinhole size is too large. I also looked at the users guide, found this formula r=0.61*lamda*N/NA. According to it, I got 238nm. Could you tell me what I should choose?
The pinhole measure as used by Huygens is the "backprojected pinhole RADIUS", which means that the physical size should by divided by the systems magnification factor. Using a 100x objective lens the value is 410 nm, which is still pretty high (250 nm is a common value), so may be there is some more internal magnification in the system we are not aware off. <br>
Using the Airy-disk measure prevents that you need to know magnification settings so the 238 nm is the one to use.
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Keywords: Leica ps2 airy disk pinhole<br>
Categories: Faq Microscopy, Huygens Faq, Imported Faqs<br>
Platforms: Irix Linux Windows Mac<br>
Related products: Hu Ess Hu Pro<br>
