How do I compute the correct sampling for 2-Photon imaging?
I am processing a 2-Photon widefield stack. I assumed that the axial resolution of the 2-Photon mic was not too good so that 500 nm sampling was probably ok for the following image: NA : 0.9; Excitation: 790 nm (2-Photon!!!); Emission: 400-500nm (Indo-1); Mounting Medium: Water: 1.30.
To find out the ideal sampling, you can always use the online Nyquist Calculator, entering the image parameters (including the number of photons: 2).
You can also use the Huygens Software in your computer. To find out how large are your samples in relation with the ideal Nyquist Rate, do the following:
In Essential:
- If you have an image for which you want to compute the Nyquist rate, open it and check its parameters (right click -> "Show parameters").
- Whenever you change the Microscopic Parameters of an image, the Nyquist rate is recalculated. Modify the image parameters to match the microscopic conditions for which you want to compute the Nyquist rate, then check the parameters again (right click -> "Show parameters").
In the Pro:
- Select the image, select Edit -> Nyquist rate in the main menu. You'll get a popup displaying the Nyquist rate.
See also Multi Photon Microscope.
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Keywords: sampling 2-photon<br>
Categories: Faq Deconvolution, Faq Microscopy, Huygens Faq, Imported Faqs<br>
Platforms: Irix Linux<br>
Related products: Hu Pro<br>
