I’m using spinning disk confocal of YOKOGAWA to capture 3D live imaging of tissue. I want to calculate the optimal resolution of Z. What kind of equation should I use for that?
- My Objective magnification: 63x and Objective N.A: 1.3, it says pinhole FWHM(z) eq.5(um) is 1.45um from the table. Does it mean my maximum resolution is 1.45um?
FWHM axial = [(0.88 × λ Em / (n - (n 2 - NA2)^1/2))2+ ((2^1/2)× n × PH / NA)2]^1/2
5th equation of this site ZEISS Microscopy Online Campus
Then, what should be the correct interval of z stack? Should the maximum resolution(1.45um) be divided by 2 or 3? why?
Even if I measure the resolution of z-stack using beads experimentally, why should I divide the value with 2 or 3?