We present a flexible, compact, 2photon, 3photon, SHG and CARS nonlinear endo-microscope featuring a 2.2 mm outer diameter and a length of 34 mm. It uses a negative curvature hollow-core double-clad fiber that is scanned with a resonant piezo-scanner. 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 >300 microns field of view is made possible using micro-lenses or GRIN based miniature objective lens. We demonstrate 2-photon and 3-photon fluorescence, SHG, THG and CARS imaging at a rate of 10 frames/s.
The lensless endoscope represents the ultimate limit in miniaturization of imaging tools: an image can be transmitted through a (multi-mode or multi-core) fiber by numerical or physical inversion of the fiber's pre- measured transmission matrix. However, the transmission matrix changes completely with only minute conformational changes of the fiber, which has so far limited lensless endoscopes to fibers that must be kept static. In this work we report a lensless endoscope which is exempt from the requirement of static fiber by designing and employing a custom-designed conformationally invariant fiber. We give experimental and theoretical validations and determine the parameter space over which the invariance is maintained.
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.
Multicore fiber bundles are attractive candidates for lensless endscopes because of their ruggedness and the relative simplicity involved in their calibration and operation. Nevertheless, the measurement of the transmission matrix of the MCF is still an interferometric process, typically requiring high stability and possibly sequential measurements. A key challenge is to replace this with a much simpler and robust technique – phase retrieval. In this talk, we will examine the major challenge in using phase retrieval in conventional MCFs. This is related to the discrete and periodic nature of the auto-correlation of an ordered MCF, resulting in the stagnation of phase retrieval algorithms in one of a multitude of local minimums. We employ phase diversity, i.e. more the one complex illumination pattern with a known phase profile can help overcome this issue. In particular, we identify that spiral phase patterns are well-suited due to their generation of complementary speckle patterns, resulting in highly non-redundant information. We experimentally demonstrate that three intensity images are sufficient to retrieve the transmission matrix with very high accuracy and success rates. Furthermore, we will also present a novel disordered MCF, which facilitates phase retrieval with a single intensity image and a priori knowledge of the core positions. This is a simple and rapidly converging method which relies on the aperiodic arrangement and the sparsity of the cores. Both these computationally inexpensive techniques highlight the potential of phase retrieval as a tool for robust phase calibration of fiber bundles in lensless imaging.
Lensless endoscopes have generated a great deal of interest in the development of minimally invasive probes for imaging in sensitive and hitherto inaccessible regions as found in deep brain imaging. Lifting the requirement for the opto-mechanical elements at the distal end of the fiber reduces the footprint of the endoscope down to the fundamental limit, the fiber itself. Our approach, using specially designed multicore fibers allows us to i) generate two-photon fluorescence contrast with femtosecond pulses, ii) image at high speeds with resonant scanners, iii) simple and non-interferometric calibration schemes and iii) exhibits a high resilience to spatio-temporal distortion of the focus due to fiber bending. In this contribution, we will discuss how novel designs of the MCFs can lift several of the instrumental complexity typically associated wavefront shaping and high speed imaging. We show that the use of sparse arrays of fiber cores can provide pixelation-free imaging with no artifacts when employed in the wavefront domain as opposed to conventional fiber bundles. Furthermore, we examine the unique properties of the MCFs which allow for fast and non-interferometric calibration schemes and can tolerate severe bending with an intact focus. The inclusion of a secondary cladding on the MCF allows us high sensitivity detection though the fiber (NA = 0.6) whilst preserving the advantages of a sparse MCF. The combination of these new developments brings us towards the application of these ultrathin probes in realistic imaging conditions.
A class of miniaturized imaged systems based on multicore fibers (MCFs) and wavefront shaping, also known as lensless endoscopes, have emerged as promising candidates for non-invasive imaging deep inside the tissue. At the current stage of their development, these systems already provide features like pixelation-free wide-field and point-scanning imaging and are compatible with the majority of nonlinear imaging techniques, offering diffraction-limited resolution over a field of view only limited by the single fiber numerical aperture.
Widely spaced single-mode cores in such an MCF are designed for very weak inter-core coupling, which ensures an infinite memory effect and provides good ultrashort pulse delivery framework for nonlinear imaging. In a true endoscopic setting, however, where the fiber geometry is subject to continuous deformation (at a rate from several Hz to several tens of Hz), results in inter-core phase and group delay dispersion (GDD) that changes as the fiber bends. This consequently degrades the PSF in terms of size and power in the focal spot, eventually rendering impossible to produce non-linear imaging.
We addressed this issue by implementing an active measurement and compensation loop at the proximal end of an MCF, allowing to correct in real time the GDD changes and to minimize the temporal dispersion of the delivered laser pulses. We evaluate this approach against a passive scheme where a static pre-compensation is used in a compliment with a specifically designed MCF, allowing to drastically increase the MCF bending robustness.
The periodic arrangement of core positions in multi-core fiber bundles introduces ‘ghost’ artifacts to endoscopic images obtained through them, whether in wide-field imaging (based on either direct imaging or speckle correlations) or in confocal scanning microscopy using wavefront shaping. Here we introduce partially disordered multi-core bundles as a means to overcome these artifacts. The benefits of their use will be discussed in the context of multiphoton scanning microscopy utilizing a spatial light modulator in the proximal end, and in the more general case of widefield imaging. We also show that both numerically and experimentally that the presence of disorder also enables to apply phase retrieval methods to characterize the phase distortion introduced due to propagation in the bundle without the need of an interferometrically stabilized reference. Thus, in addition to overcoming the challenge of ghost artifacts, disordered multi-core fibers have the potential to overcome another challenge, movement-induced phase distortions, by enabling real-time characterization of this phase distortion in reflection mode only via the proximal end.
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.
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