Abstract: The invention provides a technique for accurate detection of objects appearing in the image, such as single fluorescent molecules, cells, microorganisms, nucleotides, DNA strands, or stars in celestial images. The method allows to differentiate between two combined point sources even if they are closer to each other than the optical resolution of the system. The procedure involves computing several coefficients representing decomposition of the intensity data in the basis of Hermit functions.

Abstract: The invention provides a technique for accurate detection of objects appearing in the image, such as single fluorescent molecules, cells, microorganisms, nucleotides, DNA strands, or stars in celestial images. The method allows to differentiate between two combined point sources even if they are closer to each other than the optical resolution of the system. The procedure involves computing several coefficients representing decomposition of the intensity data in the basis of Hermit functions. In the case when the objects are distributed randomly and higher yield of accurately detected objects is desired, the method allows for a tenfold increase in such yield.

Abstract: Images with closely spaced objects can be processed using a deblending procedure that includes the calculation of some moments and centroids of intensity data. Methods and apparatus for performing this processing are well-suited for use in DNA sequencing, where the locations of fluorescing nucleotides appearing in images must be compared across several images and can be very close to one another in any single image. The increased accuracy and resolution provided by embodiments of the invention reveals previously undetected or misdetected fluorescing nucleotides, thereby facilitating the sequencing process. Embodiments of the invention can be used in other applications where, for example, defects in testing apparatus and/or limitations on image resolution frustrate subsequent analyses.