LIN S.-EJHENG D.-YHSU K.-YLIU Y.-RWEI-HSIANG HUANGHsiang-Chieh Lee2022-04-252022-04-25202121567085https://www.scopus.com/inward/record.uri?eid=2-s2.0-85110937164&doi=10.1364%2fBOE.431586&partnerID=40&md5=e29b6923f722a0e24023e1b286c303a6https://scholars.lib.ntu.edu.tw/handle/123456789/605993A technique using Linnik-based optical coherence microscopy (OCM), with built-in fluorescence microscopy (FM), is demonstrated here to describe cellular-level morphology for fresh porcine and biobank tissue specimens. The proposed method utilizes color-coding to generate digital pseudo-H&E (p-H&E) images. Using the same camera, colocalized FM images are merged with corresponding morphological OCM images using a 24-bit RGB composition process to generate position-matched p-H&E images. From receipt of dissected fresh tissue piece to generation of stitched images, the total processing time is <15 min for a 1-cm2 specimen, which is on average two times faster than frozen-section H&E process for fatty or water-rich fresh tissue specimens. This technique was successfully used to scan human and animal fresh tissue pieces, demonstrating its applicability for both biobank and veterinary purposes. We provide an in-depth comparison between p-H&E and human frozen-section H&E images acquired from the same metastatic sentinel lymph node slice (?10 μm thick), and show the differences, like elastic fibers of a tiny blood vessel and cytoplasm of tumor cells. This optical sectioning technique provides histopathologists with a convenient assessment method that outputs large-field H&E-like images of fresh tissue pieces without requiring any physical embedment. ? 2021 Optical Society of America.Blood vesselsCytologyFluorescenceFluorescence microscopyImage codingCellular levelsFrozen sectionsMicroscopic imagingOptical coherence microscopyOptical sectioningSentinel lymph nodesTissue specimensTotal processing timeTissueanimal tissueArticleconfocal microscopycontrolled studyfluorescence microscopyhumanhuman tissuenonhumanRNA extractionsentinel lymph node[SDGs]SDG3[SDGs]SDG6journal article10.1364/BOE.4315862-s2.0-85110937164