Multi-modal imaging of histological tissue sections

Two common imaging modalities for histological sections are brightfield and fluorescence microscopy imaging. Hematoxylin-Eosin (H&E) based brightfield microscopy has been the traditional imaging technique for imaging morphology, while an epi-fluorescent microscope is used for immunofluorescent staining of specific proteins or fluorescent in situ hybridization (FISH) for genetic based analysis of DNA. Simultaneous imaging of both microscopy modalities has been difficult due to optical and chemical effects of the H&E dyes. We present a novel sequential imaging and registration technique that enables brightfield and fluorescent imaging on the same tissue section, hence combining the traditional anatomic pathology with the newly emerging field of molecular pathology. First the tissue is labeled with fluorescent biomarkers, and imaged through a fluorescence microscope, and then the tissue is re-labeled with H&E dyes, and imaged again with traditional brightfield. Our robust registration algorithms achieve 99.8% registration success rate on tissue micro array (TMA) sections.