Coherent Raman scattering microscopy is a fast,label-free,and chemically specific imaging technique that shows high potential for future in vivo optical histology.However,the imaging depth in tissues is limited to the...Coherent Raman scattering microscopy is a fast,label-free,and chemically specific imaging technique that shows high potential for future in vivo optical histology.However,the imaging depth in tissues is limited to the sub-millimeter range because of absorption and scattering.Realization of coherent Raman imaging using a fiber endoscope system is a crucial step towards imaging deep inside living tissues and providing information that is inaccessible with current microscopy tools.Until now,the development of coherent Raman endoscopy has been hampered by several issues,mainly related to the fiber delivery of the excitation pulses and signal collection.Here,we present a flexible,compact,coherent Raman,and multimodal nonlinear endoscope(4.2mm outer diameter,71mm rigid length)based on a resonantly scanned hollow-core Kagomé-lattice double-clad fiber.The fiber design enables distortion-less,background-free delivery of femtosecond excitation pulses and back-collection of nonlinear signals through the same fiber.Sub-micrometer spatial resolution over a large field of view is obtained by combination of a miniature objective lens with a silica microsphere lens inserted into the fiber core.We demonstrate high-resolution,high-contrast coherent anti-Stokes Raman scattering,and second harmonic generation endoscopic imaging of biological tissues over a field of view of 320μm at a rate of 0.8 frames per second.These results pave the way for intraoperative label-free imaging applied to real-time histopathology diagnosis and surgery guidance.展开更多
基金supported by EU-ITN-607842-2013-FINON,FR-“Investissement d’Avenir”-11-IDEX-0001-02,11-INSB-0006,11-EQPX-0017,11-LABX-0007,FR-ANR-14-CE17-0004-01,FR-INSERM-PC201508,EU Regional Development Fund(ERDF)Centre National de la Recherche Scientifique(IRCICA USR 3380)the Chinese Science Council(China)for funding support.
文摘Coherent Raman scattering microscopy is a fast,label-free,and chemically specific imaging technique that shows high potential for future in vivo optical histology.However,the imaging depth in tissues is limited to the sub-millimeter range because of absorption and scattering.Realization of coherent Raman imaging using a fiber endoscope system is a crucial step towards imaging deep inside living tissues and providing information that is inaccessible with current microscopy tools.Until now,the development of coherent Raman endoscopy has been hampered by several issues,mainly related to the fiber delivery of the excitation pulses and signal collection.Here,we present a flexible,compact,coherent Raman,and multimodal nonlinear endoscope(4.2mm outer diameter,71mm rigid length)based on a resonantly scanned hollow-core Kagomé-lattice double-clad fiber.The fiber design enables distortion-less,background-free delivery of femtosecond excitation pulses and back-collection of nonlinear signals through the same fiber.Sub-micrometer spatial resolution over a large field of view is obtained by combination of a miniature objective lens with a silica microsphere lens inserted into the fiber core.We demonstrate high-resolution,high-contrast coherent anti-Stokes Raman scattering,and second harmonic generation endoscopic imaging of biological tissues over a field of view of 320μm at a rate of 0.8 frames per second.These results pave the way for intraoperative label-free imaging applied to real-time histopathology diagnosis and surgery guidance.
