Quantitative, multispectral image analysis of tissue specimens stained with quantum dots

摘要

The present invention relates to a method of whole slide imaging of a biological specimen, the specimen having 1 ... N fluorophores present in the specimen, at least one of which is a quantum dot, comprising the steps of:
(a) obtaining one or more images of the specimen with a digital camera at a low resolution;
(b) morphologically processing the one or more images or an image derived therefrom and automatically identifying one or more regions of interest containing biological structures present in the specimen;
(c) automatically obtaining an image cube of the one or more regions of interest with the camera at a high resolution, the image cube comprising image data for the regions of interest at M different wavelengths, where M is an integer greater than 2;
(d) determining from the image cube coefficients C 1 ... C N for each pixel imaging the one or more regions of interest, wherein the coefficients C 1 ... C N are related to the concentrations of the 1 ... N fluorophores present in the specimen imaged by each pixel;
(e) conducting a quantitative analysis of the regions of interest including calculating fluorophore concentrations for the biological structures from the coefficients C 1 ... C N ; and
(f) generating display data of the results of the quantitative analysis for display of a workstation.