Multiplexing ultraviolet-excited ultrasound and autofluorescence enables slide-free and label-free intraoperative histopathology imaging
Wei Song,
Xiaobao Wang,
Yiyan Zhuang,
Yechen Wang,
Qinlian Ye,
Ya-chao Wang,
Xiaocong Yuan
Affiliations
Wei Song
Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, China
Xiaobao Wang
Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, China
Yiyan Zhuang
Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, China
Yechen Wang
Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, China
Qinlian Ye
Depart of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People’s Hospital, Shenzhen 518060, China
Ya-chao Wang
Depart of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People’s Hospital, Shenzhen 518060, China
Xiaocong Yuan
Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen 518060, China
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.
