Coupling optimized bending-insensitive multi-core fibers for lensless endoscopy.

We report a bending-insensitive multi-core fiber (MCF) for lensless endoscopy imaging with modified fiber geometry that enables optimal light coupling in and out of the individual cores. In a previously reported bending insensitive MCF (twisted MCF), the cores are twisted along the length of the MCF allowing for the development of flexible thin imaging endoscopes with potential applications in dynamic and freely moving experiments. However, for such twisted MCFs the cores are seen to have an optimum coupling angle which is proportional to their radial distance from the center of the MCF. This brings coupling complexity and potentially degrades the endoscope imaging capabilities. In this study, we demonstrate that by introducing a small section (1 cm) at two ends of the MCF, where all the cores are straight and parallel to the optical axis one can rectify the above coupling and output light issues of the twisted MCF, enabling the development of bend-insensitive lensless endoscopes.

Miniature 120-beam coherent combiner with 3D-printed optics for multicore fiber-based endoscopy.

In this Letter, we report a high-efficiency, miniaturized, ultra-fast coherent beam, combined with 3D-printed micro-optics directly on the tip of a multicore fiber bundle. The highly compact device footprint (180 µm in diameter) facilitates its incorporation into a minimally invasive ultra-thin nonlinear endoscope to perform two-photon imaging.

Line-scan compressive Raman imaging with spatiospectral encoding.

We report a line-scanning imaging modality of compressive Raman technology with a single-pixel detector. The spatial information along the illumination line is encoded onto one axis of a digital micromirror device, while spectral coding masks are applied along the orthogonal direction. We demonstrate imaging and classification of three different chemical species.

Spatial frequency modulated imaging in coherent anti-Stokes Raman microscopy.

For sparse samples or in the presence of ambient light, the signal-to-noise ratio (SNR) performance of single-point-scanning coherent anti-Stokes Raman scattering (CARS) images is not optimized. As an improvement, we propose replacing the conventional CARS focus-point illumination with a periodically structured focus line while continuing to collect the transmitted CARS intensity on a single detector. The object information along the illuminated line is obtained by numerically processing the CARS signal recorded for various periods of the structured focus line. We demonstrate experimentally the feasibility of this spatial frequency modulated imaging (SPIFI) in CARS (SPIFI-CARS) and SHG (SPIFI-SHG) and identify situations where its SNR is better than that of the single-point-scanning approach.

Supercritical angle fluorescence for enhanced axial sectioning in STED microscopy.

We demonstrate subwavelength axial sectioning on biological samples with a stimulated emission depletion (STED) microscope combined with supercritical angle fluorescence (SAF) detection. SAF imaging is a powerful technique for imaging the membrane of the cell based on the direct exploitation of the fluorophore emission properties. Indeed, only when fluorophores are close to the interface can their evanescent near-field emission become propagative and be detected beyond the critical angle. Therefore, filtering out the SAF emission from the undercritical angle fluorescence (UAF) emission in the back focal plane of a high-NA objective lens permits nanometer axial sectioning of fluorescent emitters close to the coverslip. When combined with STED microscopy, a straightforward gain in axial resolution can be reached without any alteration of the STED beam path. Indeed, STED-SAF implementation only requires a modification in the detection path of the STED microscope and thus could be widely implemented.

Compressive Raman imaging with spatial frequency modulated illumination.

We report a line scanning imaging modality of compressive Raman technology with spatial frequency modulated illumination using a single pixel detector. We demonstrate the imaging and classification of three different chemical species at line scan rates of 40 Hz.

Single-shot noninterferometric measurement of the phase transmission matrix in multicore fibers.

A simple technique for far-field single-shot noninterferometric determination of the phase transmission matrix of a multicore fiber with over 100 cores is presented. This phase retrieval technique relies on the aperiodic arrangement of the cores.

Nonlinear imaging through a Fermat's golden spiral multicore fiber.

We report two-photon lensless imaging through a novel Fermat's golden spiral multicore fiber. This unique layout optimizes the sidelobe levels, field of view, crosstalk, group delay, and mode density to achieve a sidelobe contrast of at least 10.9 dB. We demonstrate experimentally the ability to generate and scan a focal point with femtosecond pulses and perform two-photon imaging.

A straightforward STED-background corrected fitting model for unbiased STED-FCS analyses.

Combining stimulated emission depletion and fluorescence correlation spectroscopy (STED-FCS) provides a powerful and sensitive tool for studying the molecular dynamics in live cells with high spatio-temporal resolution. STED-FCS gives access to molecular diffusion characteristic at the nanoscale occurring within short period of times. However due to the incomplete suppression of fluorescence in the STED process, the STED-FCS point spread function (PSF) deviates from a Gaussian shape and challenges the analysis of the auto-correlation curves obtained by FCS. Here, we model the effect of the incomplete fluorescence suppression in STED-FCS experiments and propose a new fitting model improving the accuracy of the diffusion times and average molecule numbers measurements. The implementation of a STED module with pulsed laser source on a commercial confocal/FCS microscope allowed us to apply the STED-background corrected model to fit the STED-FCS measurements. The experimental results are in good accordance with the theoretical analysis both for the number of molecules and the diffusion time which decrease accordingly with the STED power.

High-resolution multimodal flexible coherent Raman endoscope.

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 (4.2 mm outer diameter, 71 mm 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 320 µm 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.

Bending-induced inter-core group delays in multicore fibers.

We examine the impact of fiber bends on ultrashort pulse propagation in a 169-core multicore fiber (MCF) by numerical simulations and experimental measurements. We show that an L-shaped bend (where only one end of the MCF is fixed) induces significant changes in group delays that are a function of core position but linear along the bending axis with a slope directly proportional to the bending angle. For U- and S-shaped bends (where both ends of the MCF are fixed) the induced refractive index and group delay changes are much smaller than the residual, intrinsic inter-core group delay differences of the unbent MCF. We further show that when used for point-scanning lensless endoscopy with ultrashort pulse excitation, bend-induced group delays in the MCF degrade the point-spread function due to spatiotemporal coupling. Our results show that bend-induced effects in MCFs can be parametrized with only two parameters: the angle of the bend axis and the amplitude of the bend. This remains valid for bend amplitudes up to at least 200 degrees.

Phase retrieval in multicore fiber bundles.

Multicore fiber bundles are widely used in endoscopy due to their miniature size and their direct imaging capabilities. They have recently been used, in combination with spatial light modulators, in various realizations of endoscopy with little or no optics at the distal end. These schemes require characterization of the relative phase offsets between the different cores, typically done using off-axis holography, thus requiring both an interferometric setup and, typically, access to the distal tip. Here we explore the possibility of employing phase retrieval to extract the necessary phase information. We show that in the noise-free case, disordered fiber bundles are superior for phase retrieval over their periodic counterparts, and demonstrate experimentally accurate retrieval of phase information for up to 10 simultaneously illuminated cores. Thus, phase retrieval is presented as a viable alternative for real-time monitoring of phase distortions in multicore fiber bundles.

Widefield lensless endoscopy with a multicore fiber.

We demonstrate pixelation-free real-time widefield endoscopic imaging through an aperiodic multicore fiber (MCF) without any distal opto-mechanical elements or proximal scanners. Exploiting the memory effect in MCFs, the images in our system are directly obtained without any post-processing using a static wavefront correction obtained from a single calibration procedure. Our approach allows for video-rate 3D widefield imaging of incoherently illuminated objects with imaging speed not limited by the wavefront-shaping device refresh rate.

Ultrathin endoscopes based on multicore fibers and adaptive optics: a status review and perspectives.

We take stock of the progress that has been made into developing ultrathin endoscopes assisted by wave front shaping. We focus our review on multicore fiber-based lensless endoscopes intended for multiphoton imaging applications. We put the work into perspective by comparing with alternative approaches and by outlining the challenges that lie ahead.

Extended field-of-view in a lensless endoscope using an aperiodic multicore fiber.

We investigate lensless endoscopy using coherent beam combining and aperiodic multicore fibers (MCF). We show that diffracted orders, inherent to MCF with periodically arranged cores, dramatically reduce the field-of-view (FoV), and that randomness in MCF core positions can increase the FoV up to the diffraction limit set by a single fiber core, while maintaining a MCF experimental feasibility. We demonstrate experimentally pixelation-free lensless endoscopy imaging over a 120 µm FoV with an aperiodic MCF designed with widely spaced cores. We show that this system is suitable to perform beam scanning imaging by simply applying a tilt to the proximal wavefront.

Single-shot polarimetry imaging of multicore fiber.

We report an experimental test of single-shot polarimetry applied to the problem of real-time monitoring of the output polarization states in each core within a multicore fiber bundle. The technique uses a stress-engineered optical element, together with an analyzer, and provides a point spread function whose shape unambiguously reveals the polarization state of a point source. We implement this technique to monitor, simultaneously and in real time, the output polarization states of up to 180 single-mode fiber cores in both conventional and polarization-maintaining fiber bundles. We demonstrate also that the technique can be used to fully characterize the polarization properties of each individual fiber core, including eigen-polarization states, phase delay, and diattenuation.

Ultra-thin rigid endoscope: two-photon imaging through a graded-index multi-mode fiber.

Rigid endoscopes like graded-index (GRIN) lenses are known tools in biological imaging, but it is conceptually difficult to miniaturize them. In this letter, we demonstrate an ultra-thin rigid endoscope with a diameter of only 125 µm. In addition, we identify a domain where two-photon endoscopic imaging with fs-pulse excitation is possible. We validate the ultra-thin rigid endoscope consisting of a few cm of graded-index multi-mode fiber by using it to acquire optically sectioned two-photon fluorescence endoscopic images of three-dimensional samples.

Confocal supercritical angle microscopy for cell membrane imaging.

We demonstrate subwavelength sectioning on biological samples with a conventional confocal microscope. This optical sectioning is achieved by the phenomenon of supercritical angle fluorescence, wherein only a fluorophore next to the interface of a refractive index discontinuity can emit propagating components of radiation into the so-called forbidden angles. The simplicity of this technique allows it to be integrated with a high numerical aperture confocal scanning microscope by only a simple modification on the detection channel. Confocal-supercritical angular fluorescence microscopy would be a powerful tool to achieve high-resolution surface imaging, especially for membrane imaging in biological samples.