We present a flexible, compact, and multimodal nonlinear endoscope (2.2 mm outer diameter) based on a resonantly scanned negative curvature hollow-core double-clad fiber. The fiber design allows distortion-less, background-free delivery of femtosecond and picosecond excitation pulses and the back-collection of nonlinear signals through the same fiber. Sub-micron spatial resolution together with large field of view is made possible by the combination of a miniature objective lens together with a silica microsphere lens inserted into the fiber core. We demonstrate coherent anti-Stokes Raman scattering, 2-photon fluorescence and second harmonic generation imaging at a rate of 10 frames/s.
Non-linear optical microscopy proves to be an indispensable tool in natural sciences and becomes more and more attractive for clinical applications. Coherent Raman scattering, for instance, has the potential to become an in-vivo fast label-free histology tool as its chemical selectivity provides quantitative information on lipids and proteins locations and concentrations in tissues. Along with these techniques, second-harmonic generation of collagen and 2-photon excitation fluorescence broaden even more the non-linear imaging ability as collagen fibers represent an important role in human body construction. Whilst 2-photon excitation fluorescence allows to study auto-fluorescence (ex. NADH and NADHP molecules), and to excite a vast range of chromophores. However, absorption and scattering limit significantly the nonlinear imaging depth into tissues. As a solution, we offer a flexible, compact, and multimodal nonlinear endoscope (2.2 mm outer diameter, 35 mm rigid length) based on a resonantly piezo scanned hollow-core negative curvature double-clad fiber. The fiber design allows distortion-less, background-free delivery of femtosecond excitation pulses and the back-collection of nonlinear signals through the same fiber. The double-cladding of this fiber attends 10^5μm of silica collection surface which allows for a 4-fold collection improvement compared to previously used Kagomé hollow core fibers. Having a good control on the resonantly scanning fiber the endoscope can perform nonlinear imaging up to 8 frames per second over a field of view of 400μm. We demonstrate 2photon, SHG and CARS imaging in ex vivo gastric human tissue samples and in-vivo 2-photon fluorescence imaging of GFP-labeled neurons in mouse brain.
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