Sub-100-nanometre resolution in total internal reflection fluorescence microscopy

Combining total internal reflection fluorescence microscopy with structured illumination allows optical wide-field imaging with sub-100-nanometre resolution. We present a novel objective-launch set-up for standing wave illumination that takes advantage of a tunable transmission diffraction grating and transparent phase shifters actuated by electro-active polymers to control the excitation pattern in three dimensions. Image acquisition is completed in less than 1 s. To reconstruct the extended image spectrum, we apply a new apodization function that results in a lateral resolution of 89 nm for green emission wavelength.