Multi-modal contrast of tissue anatomy enables correlative biomarker imaging

Optical imaging techniques are being developed that promise to increase the information content related to specific molecular reporters. Such modalities do not produce contrast in the structural context of the surrounding tissue, making it difficult to reconcile molecular information with morphological context. We report a solution that enables visualization of the tissue morphology on formalin-fixed, paraffin embedded sections prepared for analytical biomarker imaging. Our approach combines modes of transmitted darkfield and fluorescence contrast and computer visualization to produce 2-component image data analogous to the classical hematoxylin and eosin histological stain. An interferometric hyperspectral image capture mode enables measurement of multiplexed biomarkers in annotated anatomic regions. The system enables practical correlative analysis of molecular changes within areas of anatomic pathology.