Descriptive measurements and quantification of staining artifacts for in situ hybridization

摘要

Immunohistochemistry (IHC) and in situ hybridization (ISH) have the aim of detecting, localizing and quantifying certain analytes for various diagnostic purposes. The quality of the stains which are analyzed may deviate for various reasons. Therefore, the present invention provides a method and system for assessing the stain quality and for establishing objective criteria for assessing the stain quality for application in the fields of in-situ hybridization and immunohistochemistry. In one possible embodiment, the invention comprises the steps of unmixing multi-spectral image data of a tissue specimen to obtain analyte intensity images, each analyte intensity image comprising signals from a single stain, computing metrics based on the analyte intensity images, wherein the metrics are uniformity, distribution and/or dispersion of pixel intensity values in the analyte intensity images and assessing a stain quality of a slide by comparing the computed metrics to pre-determined cutoff values regarding uniformity, distribution and/or dispersion of pixel intensity, wherein the stain quality of the slide is assessed as acceptable if the computed metrics meet or exceed the pre-determined cutoff values, and wherein the stain quality of the slide is assessed as unacceptable if the computed metrics do not meet the pre-determined cutoff values. In order to establish objective criteria for assessing stain quality, in one possible embodiment, the method and system includes the step of deriving cut-off values for uniformity, distribution and/or dispersion of pixel intensity by combining the computed metrics based on the analyte intensity images with pre-established data quantifying the stain quality.