Patent ID: 7803634
Filing Date: 2010-09-28
Classification: G01N,H04N,Y10S,Y10T

Abstract:
1. A method for nanoscopy of dyed objects being placed in a solution or as a slice of a preserved sample being placed in a polymer, wherein a fluorescent microscope having digital video-cameras with suppressing color filters for picking up fluorescence light of the object that are able to detect with a low noise level images of separate fluorescing molecules excited by a laser, the microscope having a computer for recording images received from the video-camera, the method comprising: periodically turning one to two thousands of non-fluorescing dye molecules into fluorescing fluorescing dye molecules by a UV flash, wherein the molecules cover object structures, turning the molecules into fluorescing molecules by separating chemical groups which block fluorescence, detecting images of created fluorescing molecules on video-camera frames as separate spots, wherein the molecules photobleach in the process of detection and frames with residual fluorescence are subtracted from frames containing a mixture of fluorescing particles and a residual fluorescence, saving subtracted frames to computer memory to find centers of spots which represent coordinates of dye molecules, repeating the process a number of times to calculate coordinates of all dye molecules in a lighted area of the object and reconstruct a 2-D image or a 3-D image of the object with a resolution better than 20-50 nm.