High-resolution fluorescence microscopy

摘要

A microscopy method for producing a high-resolution image of a 2-dimensional sample. The method includes exciting statistically blinking fluorophores in a sample by irradiating the sample with illumination radiation, repeatedly imaging the sample onto a spatially resolving detector in an image field that covers only a part of the sample to thereby obtain a frame sequence, generating an image from the frame sequence, the image having a spatial resolution increased beyond the optical resolution limit using a cumulant function, moving the position of the image field on the sample at least once and repeating the imaging and generating steps to obtain one image for each position of the image field, and combining the resultantly obtained images to form a complete image of the sample.

主权项

1. A microscopy method for producing a high-resolution image of a sample that contains fluorophores that, upon excitation, emit statistically blinking fluorescence radiation, the method comprising:
a) exciting the fluorophores in the sample by irradiating the sample with illumination radiation; b) repeatedly imaging the sample onto a spatially resolving detector in an image field that covers only a part of the sample, to obtain a frame sequence; c) generating an image from the frame sequence, the image having a spatial resolution increased beyond the optical resolution limit using a cumulant function; d) moving the position of the image field on the sample at least once and repeating steps b) and c) to obtain one image per position of the image field, and combining the obtained images from step d) to form a complete image of the sample.