A live oocyte is to be imaged at 520 nm through an objective of numerical aperture .
PHYSICS · Multi-part · Fermi University Physics · Optics & Photonics · Fourier optics and image formation · Fourier optics and image formation
What an Instrument Can See, Ch Seeing what the transfer function forbids: phase contrast and super-resolution
Problem
A live oocyte is to be imaged at 520 nm through an objective of numerical aperture NA=1.2.
((a)) Estimate the classical (Abbe, incoherent) resolution λ/2NA, and the resolution achievable with structured illumination, λ/4NA, stating the criterion used.
((b)) A single fluorescent label in the same specimen is detected with N=2000 photons and a point-spread-function width σPSF=110 nm. Estimate the in-principle localisation precision, and then estimate the realistic precision once background and drift are included, using the rough scaling seen in the 2006 STORM experiment described in the chapter, where a bare estimate of 1.8 nm became a measured standard deviation of 8 nm once pixel size, background and drift were included.