Dynamic conjugate F-SHARP microscopy.

Optical microscopy is an indispensable tool in biomedical sciences, but its reach in deep tissues is limited due to aberrations and scattering. This problem can be overcome by wavefront-shaping techniques, albeit at limited fields of view (FOVs). Inspired by astronomical imaging, conjugate wavefront shaping can lead to an increased field of view in microscopy, but this correction is limited to a set depth and cannot be dynamically adapted. Here, we present a conjugate wavefront-shaping scheme based on focus scanning holographic aberration probing (F-SHARP). We combine it with a compact implementation that can be readily adapted to a variety of commercial and home-built two-photon microscopes. We demonstrate the power of the method by imaging with high resolution over extended FOV (>80 µm) deeper than 400 µm inside a mouse brain through a thinned skull.

Kilohertz binary phase modulator for pulsed laser sources using a digital micromirror device.

The controlled modulation of an optical wavefront is required for aberration correction, digital phase conjugation, or patterned photostimulation. For most of these applications, it is desirable to control the wavefront modulation at the highest rates possible. The digital micromirror device (DMD) presents a cost-effective solution to achieve high-speed modulation and often exceeds the speed of the more conventional liquid crystal spatial light modulator but is inherently an amplitude modulator. Furthermore, spatial dispersion caused by DMD diffraction complicates its use with pulsed laser sources, such as those used in nonlinear microscopy. Here we introduce a DMD-based optical design that overcomes these limitations and achieves dispersion-free high-speed binary phase modulation. We show that this phase modulation can be used to switch through binary phase patterns at the rate of 20 kHz in two-photon excitation fluorescence applications.

Towards new applications using capillary waveguides.

In this paper we demonstrate the enhancement of the sensing capabilities of glass capillaries. We exploit their properties as optical and acoustic waveguides to transform them potentially into high resolution minimally invasive endoscopic devices. We show two possible applications of silica capillary waveguides demonstrating fluorescence and optical-resolution photoacoustic imaging using a single 330 µm-thick silica capillary. A nanosecond pulsed laser is focused and scanned in front of a capillary by digital phase conjugation through the silica annular ring of the capillary, used as an optical waveguide. We demonstrate optical-resolution photoacoustic images of a 30 µm-thick nylon thread using the water-filled core of the same capillary as an acoustic waveguide, resulting in a fully passive endoscopic device. Moreover, fluorescence images of 1.5 µm beads are obtained collecting the fluorescence signal through the optical waveguide. This kind of silica-capillary waveguide together with wavefront shaping techniques such as digital phase conjugation, paves the way to minimally invasive multi-modal endoscopy.

Isotropic inverse-problem approach for two-dimensional phase unwrapping.

We propose a new technique for two-dimensional phase unwrapping. The unwrapped phase is found as the solution of an inverse problem that consists in the minimization of an energy functional. The latter includes a weighted data fidelity term that favors sparsity in the error between the true and wrapped phase differences, as well as a regularizer based on higher-order total variation. One desirable feature of our method is its rotation invariance, which allows it to unwrap a much larger class of images compared to the state of the art. We demonstrate the effectiveness of our method through several experiments on simulated and real data obtained through the tomographic phase microscope. The proposed method can enhance the applicability and outreach of techniques that rely on quantitative phase evaluation.

Delivery of focused short pulses through a multimode fiber.

Light propagation through multimode fibers suffers from spatial distortions that lead to a scrambled intensity profile. In previous work, the correction of such distortions using various wavefront control methods has been demonstrated in the continuous wave case. However, in the ultra-fast pulse regime, modal dispersion temporally broadens a pulse after propagation. Here, we present a method that compensates for spatial distortions and mitigates temporal broadening due to modal dispersion by a selective phase conjugation process in which only modes of similar group velocities are excited. The selectively excited modes are forced to follow certain paths through the multimode fiber and interfere constructively at the distal tip to form a focused spot with minimal temporal broadening. We demonstrate the delivery of focused 500 fs pulses through a 30 cm long step-index multimode fiber. The achieved pulse duration corresponds to approximately 1/30th of the duration obtained if modal dispersion was not controlled. Moreover, we measured a detailed two-dimensional map of the pulse duration at the output of the fiber and confirmed that the focused spot produces a two-photon absorption effect. This work opens new possibilities for ultra-thin multiphoton imaging through multimode fibers.

Dynamic bending compensation while focusing through a multimode fiber.

Multimode fiber endoscopes have recently been shown to provide sub-micrometer resolution, however, imaging through a multimode fiber is highly sensitive to bending. Here we describe the implementation of a coherent beacon source placed at the distal tip of the multimode fiber, which can be used to compensate for the effects of bending. In the first part of this paper, we show that a diffraction limited focused spot can be generated at the distal tip of the multimode fiber using the beacon. In the second part, we demonstrate focusing even when the fiber is bent by dynamically compensating for it. The speckle pattern at the proximal fiber end, generated by the beacon source placed at its distal end, is highly dependent on the fiber conformation. We experimentally show that by intensity correlation, it is possible to identify the fiber conformation and maintain a focus spot while the fiber is bent over a certain range. Once the fiber configuration is determined, previously calibrated phase patterns could be stored for each fiber conformation and used to scan the distal spot and perform imaging.

Increasing the imaging capabilities of multimode fibers by exploiting the properties of highly scattering media.

We present a design that exploits the focusing properties of scattering media to increase the resolution and the working distance of multimode fiber (MMF)-based imaging devices. Placing a highly scattering medium in front of the distal tip of the MMF enables the formation of smaller sized foci at increased working distances away from the fiber tip. We perform a parametric study of the effect of the working distance and the separation between the fiber and the scattering medium on the focus size. We experimentally demonstrate submicrometer focused spots as far away as 800 µm with 532 nm light.

High-resolution, lensless endoscope based on digital scanning through a multimode optical fiber.

We propose and experimentally demonstrate an ultra-thin rigid endoscope (450 µm diameter) based on a passive multimode optical fiber. We use digital phase conjugation to overcome the modal scrambling of the fiber to tightly focus and scan the laser light at its distal end. By exploiting the maximum number of modes available, sub-micron resolution, high quality fluorescence images of neuronal cells were acquired. The imaging system is evaluated in terms of fluorescence collection efficiency, resolution and field of view. The small diameter of the proposed endoscope, along with its high quality images offer an opportunity for minimally invasive medical endoscopic imaging and diagnosis based on cellular phenotype via direct tissue penetration.

Focusing and scanning light through a multimode optical fiber using digital phase conjugation.

We demonstrate for the first time to our knowledge a digital phase conjugation technique for generating a sharp focus point at the end of a multimode optical fiber. A sharp focus with a contrast of 1800 is experimentally obtained at the tip of a 105 µm core multimode fiber. Scanning of the focal point is also demonstrated by digital means. Effects from illumination and fiber bending are addressed.