Amyloid proteins are associated with a broad spectrum of neurodegenerative diseases.However,it remains a grand challenge to extract molecular structure information from intracellular amyloid proteins in their native c...Amyloid proteins are associated with a broad spectrum of neurodegenerative diseases.However,it remains a grand challenge to extract molecular structure information from intracellular amyloid proteins in their native cellular environment.To address this challenge,we developed a computational chemical microscope integrating 3D midinfrared photothermal imaging with fluorescence imaging,termed Fluorescence-guided Bond-Selective Intensity Diffraction Tomography(FBS-IDT).Based on a low-cost and simple optical design,FBS-IDT enables chemical-specific volumetric imaging and 3D site-specific mid-IR fingerprint spectroscopic analysis of tau fbrils,an important type of amyloid protein aggregates,in their intracellular environment.Label-free volumetric chemical imaging of human cells with/without seeded tau fibrils is demonstrated to show the potential correlation between lipid accumulation and tau aggregate formation.Depth-resolved mid-infrared fingerprint spectroscopy is performed to reveal the protein secondary structure of the intracellular tau fibrils.3D visualization of theβ-sheet for tau fibril structure is achieved.展开更多
Recent years have witnessed the tremendous development of fusing fiber-optic imaging with supervised deep learning to enable high-quality imaging of hard-to-reach areas.Nevertheless,the supervised deep learning method...Recent years have witnessed the tremendous development of fusing fiber-optic imaging with supervised deep learning to enable high-quality imaging of hard-to-reach areas.Nevertheless,the supervised deep learning method imposes strict constraints on fiber-optic imaging systems,where the input objects and the fiber outputs have to be collected in pairs.To unleash the full potential of fiber-optic imaging,unsupervised image reconstruction is in demand.Unfortunately,neither optical fiber bundles nor multimode fibers can achieve a point-to-point transmission of the object with a high sampling density,as is a prerequisite for unsupervised image reconstruction.The recently proposed disordered fibers offer a new solution based on the transverse Anderson localization.Here,we demonstrate unsupervised full-color imaging with a cellular resolution through a meter-long disordered fiber in both transmission and reflection modes.The unsupervised image reconstruction consists of two stages.In the first stage,we perform a pixel-wise standardization on the fiber outputs using the statistics of the objects.In the second stage,we recover the fine details of the reconstructions through a generative adversarial network.Unsupervised image reconstruction does not need paired images,enabling a much more flexible calibration under various conditions.Our new solution achieves full-color high-fidelity cell imaging within a working distance of at least 4 mm by only collecting the fiber outputs after an initial calibration.High imaging robustness is also demonstrated when the disordered fiber is bent with a central angle of 60°.Moreover,the cross-domain generality on unseen objects is shown to be enhanced with a diversified object set.展开更多
基金supported by the National Institute of General Medical Sciences(R35GM136223)a grant from Daylight Solutions,and a grant(2023-321163)the Chan Zuckerberg Initiative Donor-Advised Fund at the Silicon Valley Community Foundation.
文摘Amyloid proteins are associated with a broad spectrum of neurodegenerative diseases.However,it remains a grand challenge to extract molecular structure information from intracellular amyloid proteins in their native cellular environment.To address this challenge,we developed a computational chemical microscope integrating 3D midinfrared photothermal imaging with fluorescence imaging,termed Fluorescence-guided Bond-Selective Intensity Diffraction Tomography(FBS-IDT).Based on a low-cost and simple optical design,FBS-IDT enables chemical-specific volumetric imaging and 3D site-specific mid-IR fingerprint spectroscopic analysis of tau fbrils,an important type of amyloid protein aggregates,in their intracellular environment.Label-free volumetric chemical imaging of human cells with/without seeded tau fibrils is demonstrated to show the potential correlation between lipid accumulation and tau aggregate formation.Depth-resolved mid-infrared fingerprint spectroscopy is performed to reveal the protein secondary structure of the intracellular tau fibrils.3D visualization of theβ-sheet for tau fibril structure is achieved.
基金The authors would like to thank the valuable discussions provided by the Fiber Optics Lab at CREOL.
文摘Recent years have witnessed the tremendous development of fusing fiber-optic imaging with supervised deep learning to enable high-quality imaging of hard-to-reach areas.Nevertheless,the supervised deep learning method imposes strict constraints on fiber-optic imaging systems,where the input objects and the fiber outputs have to be collected in pairs.To unleash the full potential of fiber-optic imaging,unsupervised image reconstruction is in demand.Unfortunately,neither optical fiber bundles nor multimode fibers can achieve a point-to-point transmission of the object with a high sampling density,as is a prerequisite for unsupervised image reconstruction.The recently proposed disordered fibers offer a new solution based on the transverse Anderson localization.Here,we demonstrate unsupervised full-color imaging with a cellular resolution through a meter-long disordered fiber in both transmission and reflection modes.The unsupervised image reconstruction consists of two stages.In the first stage,we perform a pixel-wise standardization on the fiber outputs using the statistics of the objects.In the second stage,we recover the fine details of the reconstructions through a generative adversarial network.Unsupervised image reconstruction does not need paired images,enabling a much more flexible calibration under various conditions.Our new solution achieves full-color high-fidelity cell imaging within a working distance of at least 4 mm by only collecting the fiber outputs after an initial calibration.High imaging robustness is also demonstrated when the disordered fiber is bent with a central angle of 60°.Moreover,the cross-domain generality on unseen objects is shown to be enhanced with a diversified object set.
