Histopathology relies upon the staining and sectioning of biological tissues,which can be laborious and may cause artifacts and distort tissues.We develop label-free volumetric imaging of thick-tissue slides,exploitin...Histopathology relies upon the staining and sectioning of biological tissues,which can be laborious and may cause artifacts and distort tissues.We develop label-free volumetric imaging of thick-tissue slides,exploiting refractive index distributions as intrinsic imaging contrast.The present method systematically exploits label-free quantitative phase imaging techniques,volumetric reconstruction of intrinsic refractive index distributions in tissues,and numerical algorithms for the seamless stitching of multiple three-dimensional tomograms and for reducing scattering-induced image distortion.We demonstrated label-free volumetric imaging of thick tissues with the field of view of 2 mm×1.75 mm×0.2 mm with a spatial resolution of 170 nm×170 nm×1400 nm.The number of optical modes,calculated as the reconstructed volume divided by the size of the point spread function,was∼20 giga voxels.We have also demonstrated that different tumor types and a variety of precursor lesions and pathologies can be visualized with the present method.展开更多
A major challenge in three-dimensional(3D)microscopy is to obtain accurate spatial information while simultaneously keeping the microscopic samples in their native states.In conventional 3D microscopy,axial resolution...A major challenge in three-dimensional(3D)microscopy is to obtain accurate spatial information while simultaneously keeping the microscopic samples in their native states.In conventional 3D microscopy,axial resolution is inferior to spatial resolution due to the inaccessibility to side scattering signals.In this study,we demonstrate the isotropic microtomography of free-floating samples by optically rotating a sample.Contrary to previous approaches using optical tweezers with multiple foci which are only applicable to simple shapes,we exploited 3D structured light traps that can stably rotate freestanding complex-shaped microscopic specimens,and side scattering information is measured at various sample orientations to achieve isotropic resolution.The proposed method yields an isotropic resolution of 230 nm and captures structural details of colloidal multimers and live red blood cells,which are inaccessible using conventional tomographic microscopy.We envision that the proposed approach can be deployed for solving diverse imaging problems that are beyond the examples shown here.展开更多
基金H.H.,R.H.H.,S.-M.H.,and Y.P.conceived the initial idea.H.H.developed the optical system and analysis methods.H.H.and Y.W.K.performed the experiments and analyzed the data.M.L.and S.S.provided the analysis methods and analyzed the data.All authors wrote and revised the manuscript.This work was supported by KAIST,Up Program,BK21+program,Tomocube,and National Research Foundation of Korea(2017M3C1A3013923,2015R1A3A2066550,and 2018K000396).Professor Park and Mr.Moosung Lee have financial interests in Tomocube Inc.,a company that commercializes optical diffraction tomography and quantitative phase imaging instruments and is one of the sponsors of the work.
文摘Histopathology relies upon the staining and sectioning of biological tissues,which can be laborious and may cause artifacts and distort tissues.We develop label-free volumetric imaging of thick-tissue slides,exploiting refractive index distributions as intrinsic imaging contrast.The present method systematically exploits label-free quantitative phase imaging techniques,volumetric reconstruction of intrinsic refractive index distributions in tissues,and numerical algorithms for the seamless stitching of multiple three-dimensional tomograms and for reducing scattering-induced image distortion.We demonstrated label-free volumetric imaging of thick tissues with the field of view of 2 mm×1.75 mm×0.2 mm with a spatial resolution of 170 nm×170 nm×1400 nm.The number of optical modes,calculated as the reconstructed volume divided by the size of the point spread function,was∼20 giga voxels.We have also demonstrated that different tumor types and a variety of precursor lesions and pathologies can be visualized with the present method.
基金KAIST UP programme,BK21+programme,Tomocube,and National Research Foundation of Korea(2017M3C1A3013923,2015R1A3A2066550,2018K000396).
文摘A major challenge in three-dimensional(3D)microscopy is to obtain accurate spatial information while simultaneously keeping the microscopic samples in their native states.In conventional 3D microscopy,axial resolution is inferior to spatial resolution due to the inaccessibility to side scattering signals.In this study,we demonstrate the isotropic microtomography of free-floating samples by optically rotating a sample.Contrary to previous approaches using optical tweezers with multiple foci which are only applicable to simple shapes,we exploited 3D structured light traps that can stably rotate freestanding complex-shaped microscopic specimens,and side scattering information is measured at various sample orientations to achieve isotropic resolution.The proposed method yields an isotropic resolution of 230 nm and captures structural details of colloidal multimers and live red blood cells,which are inaccessible using conventional tomographic microscopy.We envision that the proposed approach can be deployed for solving diverse imaging problems that are beyond the examples shown here.
