Method for simultaneous detection of multiple fluorophores for in situ hybridization and chromosome painting

摘要

A spectral bio-imaging method characterized by high spatial and high spectral resolutions, the method comprising the steps of: (a) preparing a sample to be spectrally imaged; (b) viewing said sample through an optical device, said optical device being optically connected to an imaging spectrometer, said optical device and said imaging spectrometer being for obtaining a spectrum of each pixel of said sample by: (i) collecting incident light simultaneously from all pixels of said sample using 710 י" ב באב התש" ס - August 13, 2000 collimating optics; (ii) passing said incident collimated light through an interferometer system having a number of elements, so that said light is first split into two coherent beams which travel in different directions inside said interferometer and then said two coherent beams recombine to interfere with each other to form an exiting light beam; (iii) passing said exiting light beam through a focusing optical system which focuses said exiting light beam on a detector having a two-dimensional array of detector elements, so that at each instant each of said detector elements is the image of one and always the same pixel of said sample for the entire duration of the measurement, so that the real image of the sample is stationary on the plane of the detector array and at any time during the measurement the image is still visible and recognizable, and so that each of said detector elements produces a signal which is a particular linear combination of light intensity emitted by said pixel at different wavelengths, wherein said linear combination is a function of the instantaneous optical path difference; (iv) translating one or more of said elements of said interferometer system, so that said optical path difference between said two coherent beams generated by said interferometer system is scanned simultaneously for all said pixels of said sample; and (v) recording signals of each of said detector elements as function of time using a recording device to form a first spectral cube of data; and (c) interpreting said first spectral cube of data using a mathematical algorithm.