Far‐field fluorescence microscopy with three‐dimensional resolution in the 100‐nm range

Abstract

We report three-dimensional (3D) microscopy with nearly isotropic resolution in the lambda/5-lambda/10 range. Our approach combines 4PI-confocal two-photon fluorescence microscopy with image restoration. The 3D resolution is demonstrated with densely clustered beads as well as with F-actin fibers in mouse fibroblast cells. A comparison with unrestored two-photon confocal images reveals a total reduction of the uncertainty volume up to a factor of 15.