Patent ID: 9575302
Date: 2017-02-21
CPC Classifications: G01N,G02B

Claim:
1. A method of stimulated emission depletion microscopy using adaptive optics comprising: (a) capturing a fluorescence image from a stimulated emission depletion microscope having an excitation light path and a first light modulator in a depletion light path defining a pattern on the first light modulator, wherein the first light modulator is a spatial light modulator or a deformable mirror; (b) calculating a measure of image sharpness of the fluorescence image, calculating a measure of image brightness of the fluorescence image, and calculating a metric combining the measure of image brightness and the measure of image sharpness; wherein the metric M is defined by wherein M is the metric, S an image sharpness measure, B an image brightness measure, σ is constant that has a value −1 or 1 depending on whether the aberration mode being corrected produces a maximum or minimum brightness, β is a constant and f(S) is a function of S that has a larger absolute value for a value of S above a threshold value S T and a lower absolute value for a value of S below the threshold value S T : (c) adjusting the pattern on the first light modulator; and repeating (a), (b) and (c) to maximise or minimise the metric combining the measure of image brightness and the measure of image sharpness.