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Nonlinear optical endomicroscopy for label-free functional histology in vivo

Light(Science & Applications) 2017年第1期6卷 424-430页

作者： Wenxuan Liang Gunnsteinn Hall bernhard messerschmidt Ming-Jun Li Xingde Li Department of Biomedical Engineering Johns Hopkins UniversityBaltimoreMD 21205USA GRINTECH GmbH Jena 07745Germany Science and Technology Division Corning IncorporatedCorningNY 14831USA

This manuscript reports on the first two-photon,label-free,metabolic imaging of biological tissues in vivo at histological resolution on an extremely compact,fiber-optic endomicroscopy *** system provides new opportunities for performing noninvasive and functional histological imaging of internal organs in vivo,in situ and in real *** a routine clinical procedure,traditional histology has made significant impacts on ***,the procedure is invasive and time consuming,suffers random sampling errors,and cannot provide in vivo functional *** technology reported here features an extremely compact and flexible fiber-optic probe~2 mm in diameter,enabling direct access to internal *** two-photon imaging quality comparable to a large bench-top laser scanning microscope was achieved through technological innovations in double-clad fiber optics and miniature objective lenses(among many others).In addition to real-time label-free visualization of biological tissues in situ with subcellular histological detail,we demonstrated for the first time in vivo two-photon endomicroscopic metabolic imaging on a functioning mouse kidney *** breakthroughs in nonlinear endoscopic imaging capability present numerous promising opportunities for paradigm-shifting applications in both clinical diagnosis and basic research.

关键词： functional histology in vivo label-free imaging nonlinear endomicroscopy two-photon metabolic imaging

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Multimodal nonlinear endomicroscopic imaging probe using a double-core double-clad fiber and focus-combining micro-optical concept

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Light(Science & Applications) 2021年第11期10卷 2173-2183页

作者： Ekaterina Pshenay-Severin Hyeonsoo Bae Karl Reichwald Gregor Matz Jörg Bierlich Jens Kobelke Adrian Lorenz Anka Schwuchow Tobias Meyer-Zedler Michael Schmitt bernhard messerschmidt Juergen Popp GRINTECH GmbH Schillerstr.107745JenaGermany Leibniz Institute of Photonic Technology Member of Leibniz Health TechnologiesAlbert-Einstein-Str.907745JenaGermany Institute of Physical Chemistry and Abbe Center of Photonics Friedrich Schiller University JenaHelmholtzweg 407743JenaGermany

Multimodal non-linear microscopy combining coherent anti-Stokes Raman scattering,second harmonic generation,and two-photon excited fluorescence has proved to be a versatile and powerful tool enabling the label-free investigation of tissue structure,molecular composition,and correlation with function and disease *** a routine medical application,the implementation of this approach into an in vivo imaging endoscope is ***,this is a difficult task due to the requirements of a multicolour ultrashort laser delivery from a compact and robust laser source through a fiber with low losses and temporal synchronization,the efficient signal collection in epi-direction,the need for small-diameter but highly corrected endomicroobjectives of high numerical aperture and compact ***,we introduce an ultra-compact fiber-scanning endoscope platform for multimodal non-linear endomicroscopy in combination with a compact four-wave mixing based fiber *** heart of this fiber-scanning endoscope is an in-house custom-designed,single mode,double clad,double core pure silica fiber in combination with a 2.4 mm diameter NIR-dual-waveband corrected endomicroscopic objective of 0.55 numerical aperture and 180µm field of view for non-linear imaging,allowing a background free,low-loss,high peak power laser delivery,and an efficient signal collection in backward direction.A linear diffractive optical grating overlays pump and Stokes laser foci across the full field of view,such that diffraction-limited performance is demonstrated for tissue imaging at one frame per second with sub-micron spatial resolution and at a high transmission of 65%from the laser to the specimen using a distal resonant fiber scanner.

关键词： fiber double scanner

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