초록 ▼

Reliable autofocus is required to obtain accurate measurements of fluorescent stained cellular components from a system capable of scanning multiple microscope fields. Autofocus could be performed directly with fluorescence images, but due to photobleaching and destructive fluorescence by-products,

Reliable autofocus is required to obtain accurate measurements of fluorescent stained cellular components from a system capable of scanning multiple microscope fields. Autofocus could be performed directly with fluorescence images, but due to photobleaching and destructive fluorescence by-products, it is best to minimize fluorescence exposure for photosensitive specimens and live cells. This exposure problem could be completely avoided by using phase-contrast microscopy, implemented through the same optics as fluorescence microscopy. Functions for both phase-contrast and fluorescence autofocus were evaluated using the present invention and the suitability of phase-contrast autofocus for fluorescence microscopy was determined. The present autofocus system for scanning microscopy can be performed at least as fast as 0.25 s/field without loss of precision.

대표청구항 ▼

[ What is claimed is:] [1.] A sequential method of autofocus for a microscope having a focus mechanism, the method comprising:scanning a specimen on a microscope stage;sequentially receiving digital images of the specimen at a plurality of vertical positions along a Z axis;for each vertical position

[ What is claimed is:] [1.] A sequential method of autofocus for a microscope having a focus mechanism, the method comprising:scanning a specimen on a microscope stage;sequentially receiving digital images of the specimen at a plurality of vertical positions along a Z axis;for each vertical position of the plurality of vertical positions:applying a digital filter to a digital image acquired at the vertical position to obtain a filtered image; andapplying a measurement function to the filtered image to obtain a value representing a degree of focus;applying a weight to the value representing a degree of focus to obtain a weighted value;applying a non-interative function to a plurality of weighted values of focus to obtain a focus position; andcausing the microscope stage to move to a vertical position corresponding to the focus position.