Patent ID: 9551658
Date: 2017-01-24
CPC Classifications: G01N,G02B

Claim:
1. A STED fluorescence light microscopic method of imaging a structure which is marked with fluorescent entities in a sample, the method comprising the steps of: applying pulses of excitation light to the sample, which excite the fluorescent entities for fluorescence, and which are focused on at least one focal area; applying de-excitation light to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave; registering fluorescence light spontaneously emitted by the excited fluorescent entities in the sample after each pulse of excitation light and overlapping with applying the de-excitation light; and repeating the steps of applying and registering at different positions of the focal area of the excitation light and the intensity zero point of the de-excitation light; wherein, in the step of registering, the fluorescence light spontaneously emitted by the excited fluorescent entities is registered with temporal resolution between consecutive pulses of the excitation light.