Rapid pseudo-H&E imaging using a fluorescence-inbuilt optical coherence microscopic imaging system
Journal
Biomedical Optics Express
Journal Volume
12
Journal Issue
8
Pages
5139-5158
Date Issued
2021
Author(s)
Abstract
A 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.
Subjects
Blood vessels
Cytology
Fluorescence
Fluorescence microscopy
Image coding
Cellular levels
Frozen sections
Microscopic imaging
Optical coherence microscopy
Optical sectioning
Sentinel lymph nodes
Tissue specimens
Total processing time
Tissue
animal tissue
Article
confocal microscopy
controlled study
fluorescence microscopy
human
human tissue
nonhuman
RNA extraction
sentinel lymph node
Type
journal article