Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging

Histological examination of tissue remains the gold standard for analysis of various diseases in both clinical diagnosis and basic research. However, long-standing challenges in conventional histology have arisen from complex sample preparations involving micrometer-thick sectioning and hematoxylin...

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Main Authors: Wei Song, Xiaobao Wang, Yiyan Zhuang, Yechen Wang, Qinlian Ye, Ya-chao Wang, Xiaocong Yuan
Format: Article
Language:English
Published: AIP Publishing LLC 2024-11-01
Series:APL Photonics
Online Access:http://dx.doi.org/10.1063/5.0232343
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author Wei Song
Xiaobao Wang
Yiyan Zhuang
Yechen Wang
Qinlian Ye
Ya-chao Wang
Xiaocong Yuan
author_facet Wei Song
Xiaobao Wang
Yiyan Zhuang
Yechen Wang
Qinlian Ye
Ya-chao Wang
Xiaocong Yuan
author_sort Wei Song
collection DOAJ
description Histological examination of tissue remains the gold standard for analysis of various diseases in both clinical diagnosis and basic research. However, long-standing challenges in conventional histology have arisen from complex sample preparations involving micrometer-thick sectioning and hematoxylin and eosin (H&E) staining processes, which lead to significant delays in accessing the diagnostic output. We propose a hybrid optical-ultrasonic microscope (HOME) that multiplexes the intrinsic thermoelastic ultrasound and fluorescence emission generated by endogenous biomolecules under single-shot ultraviolet excitation to achieve direct histopathological imaging of fresh tissue without the sectioning and staining steps. The HOME detects ultrasounds of DNA/RNA using a photoacoustic microscopy subsystem to provide a hematoxylin-like contrast that specifically highlights the cell nucleus, and it also captures the autofluorescence of endogenous fluorophores in its confocal fluorescence microscopy subsystem to offer eosin-like contrast for visualization of the cytoplasm. Parallel acquisition of photoacoustic and autofluorescence images enables comprehensive observations of co-registered cellular structures and organizations, corresponding closely to the typical histological landscapes obtained via standard H&E staining. The HOME demonstrates fast and accurate histopathological imaging of fresh murine tissue and unprocessed clinical glioma specimens and is, therefore, promising for use in assisting intraoperative pathological assessments and performing basic scientific research on neoplasms.
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institution Kabale University
issn 2378-0967
language English
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series APL Photonics
spelling doaj-art-638b93801f934f948be8fed8111d36302024-12-04T17:05:05ZengAIP Publishing LLCAPL Photonics2378-09672024-11-01911116102116102-1410.1063/5.0232343Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imagingWei Song0Xiaobao Wang1Yiyan Zhuang2Yechen Wang3Qinlian Ye4Ya-chao Wang5Xiaocong Yuan6Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, ChinaNanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, ChinaNanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, ChinaNanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, ChinaDepart of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People’s Hospital, Shenzhen 518060, ChinaDepart of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People’s Hospital, Shenzhen 518060, ChinaNanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, ChinaHistological examination of tissue remains the gold standard for analysis of various diseases in both clinical diagnosis and basic research. However, long-standing challenges in conventional histology have arisen from complex sample preparations involving micrometer-thick sectioning and hematoxylin and eosin (H&E) staining processes, which lead to significant delays in accessing the diagnostic output. We propose a hybrid optical-ultrasonic microscope (HOME) that multiplexes the intrinsic thermoelastic ultrasound and fluorescence emission generated by endogenous biomolecules under single-shot ultraviolet excitation to achieve direct histopathological imaging of fresh tissue without the sectioning and staining steps. The HOME detects ultrasounds of DNA/RNA using a photoacoustic microscopy subsystem to provide a hematoxylin-like contrast that specifically highlights the cell nucleus, and it also captures the autofluorescence of endogenous fluorophores in its confocal fluorescence microscopy subsystem to offer eosin-like contrast for visualization of the cytoplasm. Parallel acquisition of photoacoustic and autofluorescence images enables comprehensive observations of co-registered cellular structures and organizations, corresponding closely to the typical histological landscapes obtained via standard H&E staining. The HOME demonstrates fast and accurate histopathological imaging of fresh murine tissue and unprocessed clinical glioma specimens and is, therefore, promising for use in assisting intraoperative pathological assessments and performing basic scientific research on neoplasms.http://dx.doi.org/10.1063/5.0232343
spellingShingle Wei Song
Xiaobao Wang
Yiyan Zhuang
Yechen Wang
Qinlian Ye
Ya-chao Wang
Xiaocong Yuan
Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
APL Photonics
title Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
title_full Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
title_fullStr Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
title_full_unstemmed Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
title_short Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
title_sort multiplexing ultraviolet excited ultrasound and autofluorescence enables slide free and label free intraoperative histopathology imaging
url http://dx.doi.org/10.1063/5.0232343
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