Reproducible quantification of biomarker expression

摘要

A method is described for the reproducible quantification of biomarker expression, including biomarker expression in a tissue sample. Methods and systems are described whereby reproducible scores for biomarker expression are obtained independent of instrument, its location, or operator.

主权项

1. A method of reproducibly quantifying biomarker expression in a slide-mounted tissue sample, the method comprising:
(a) obtaining a slide-mounted tissue sample, which has been stained to permit localization of at least one cellular compartment and at least one biomarker; (b) obtaining one or more pixel-comprised images of the stained tissue sample using a microscope, and analyzing the one or more pixel-comprised images to obtain one or more data sets; (c) automatically analyzing the one or more data sets derived from the image pixels to differentiate data signal from noise; (d) automatically analyzing the one or more data sets derived from the image pixels to differentiate data signal attributable to each of said at least one cellular compartment; (e) optionally automatically analyzing the one or more data sets derived from the image pixels to differentiate data signal attributable to said at least one biomarker for each of said at least one cellular compartment; (f) quantifying the amount of biomarker expressed in each of said at least one cellular compartment to arrive at a standardized score, which is a product of a raw score and one or more factors selected from the group consisting of a calibration cube factor, a light source factor and an optical path factor, whereby a run comprises steps (a)-(f), and the biomarker expression in the same slide-mounted tissue sample is quantified in a manner having a level of reproducibility above 80 percent for separate runs, in which each automatic analysis step is carried out in an unsupervised manner and comprises an unsupervised pixel-based clustering algorithm, and whereby the microscope allows for automatic adjustment of exposure time to provide an optimized dynamic range of data captured in the image pixels.