Patent ID: 6438261
Filing Date: 2002-08-20
Classification: G01N,G06K

Abstract:
A method for in-situ focus-fusion multi-layer spectral imaging and analysis of a particulate sample, particles of the particulate sample exhibiting layer or depth dependent features and characteristics, the method comprising the steps of:(a) preparing the sample for the in-situ focus-fusion multi-layer spectral imaging and analysis; (b) providing a spectroscopic imaging system, including a light illumination source and a sample holder moveable by a three dimensional translation stage, for in-situ spectral imaging of the sample; (c) selecting and defining imaging scenario parameters for acquiring and analyzing in-situ spectral images of the sample, said imaging scenario parameters are particle physicochemical information and characteristics relating to particle chemical composition and particle morphology, and, particle spectral information and characteristics relating to pixel energy level or intensity, I(p), signal-to-noise ratio, (S/N), image sharpness, spectral distances, and spectral fingerprints relating to spectral emission patterns of the particles of the sample, each said spectral emission pattern featuring emission intensity as a function of wavelength of light emitted by the sample; (d) adjusting and setting said spectroscopic imaging system for said in-situ spectral imaging of the sample at a selected i-th field of view, FOVi, having central (x, y) position coordinates relative to said translation stage; (e) acquiring a cube (spectral) plane image of the sample in said selected i-th field of view, FOVi, at a selected j-th differential imaging/focusing distance, &Dgr;zij, in a selected range of imaging distance defined along z-direction between the sample and said light illumination source of said spectroscopic imaging system, by focusing said imaging system in said z-direction until receiving a sharp gray level image of the sample; (f) constructing and analyzing a focused cube (spectral) plane imageij, of the sample for said selected i-th field of view, FOVi, at said selected j-th differential imaging/focusing distance, &Dgr;zij, from said acquired cube (spectral) plane image of the sample, whereby said constructing and analyzing features identifying targets in said sharp gray level image of the sample by using said selected and said defined particle physicochemical imaging scenario parameters and said selected and said defined particle spectral imaging scenario parameters; (g) repeating step (e) and step (f) in said selected i-th field of view, FOVi, for a plurality of said selected j-th differential imaging/focusing distances, &Dgr;zij, in said selected range of imaging distance, for forming a plurality of said focused cube (spectral) plane imagesij of the sample for said selected i-th field of view, FOVi; (h) constructing a fused focused cube (spectral) imagei, for said selected i-th field of view, FOVi, using a plurality of said identified targets having high content selected from said plurality of said focused cube (spectral) plane imagesij, and, using spectral background parameters, Bi, empirically determined from said plurality of said focused cube (spectral) plane imagesij; (i) acquiring and constructing a plurality of said fused focused cube (spectral) imagesi of the sample in other said selected i-th fields of view, FOVi, having corresponding said central (x, y) position coordinates relative to said translation stage, for a plurality of said selected j-th differential imaging/focusing distances, &Dgr;zij, by repeating step (d) through step (h), until selected sample viewing/imaging range is imaged and analyzed; (j) applying at least one image analysis algorithm to data of said plurality of said fused focused cube (spectral) imagesi for identifying spectral fingerprints relating to physicochemical characterization of the sample; and (k) repeating step (d) through step (j) following each predetermined time interval, &Dgr;t, over a period of time spanning a multiple of said pre-determined time interval, &Dgr;t, for generating a statistical analysis report describing time variation of physicochemical and spectral imaging characteristics of the particulate sample.