Nicolino Stasio

Lensless two-photon imaging through a multicore fiber with coherence-gated digital phase conjugation

Abstract. We performed near-diffraction limited two-photon fluorescence (TPF) imaging through a lensless, multicore-fiber (MCF) endoscope utilizing digital phase conjugation. The phase conjugation technique is compatible with commercially available MCFs with high core density. We demonstrate focusing of ultrashort pulses through an MCF and show that the method allows for resolution that is not limited by the MCF core spacing. We constructed TPF images of fluorescent beads and cells by digital scanning of the phase-conjugated focus on the target object and collection of the emitted fluorescence through the MCF.

Resolution enhancement in nonlinear scanning microscopy through post-detection digital computation

In the last decade, the resolution of optical microscopy has been doubled thanks to linear structured illumination. The resolution has been further improved by combining structured illumination with nonlinear photoresponse. Recently, structured illumination has been combined with point-scanning microscopy. In this paper, we investigate whether, as in wide field acquisition, significant resolution enhancement can be obtained by harnessing the nonlinear response of the sample when point-scanning structured illumination is employed. We compare point scanning with wide field structured illumination microscopy in terms of signal-to-noise ratio. We conclude that superresolution using saturated point-scanning structured illumination is severely restricted to the first nonlinear orders. We identify possibilities for how different beam shapes or nonlinear phenomena might be envisaged for future implementations.

Calibration-free imaging through a multicore fiber using speckle scanning microscopy

The images produced by multicore endoscopes are pixelated, and their resolution is limited by the core-to-core spacing. Lenses can be used to improve the resolution, but this reduces the field of view proportionally. Lensless endoscopy through multicore fibers can be achieved by using wavefront shaping techniques. This requires a calibration step, and the conformation of the fiber must remain constant over time. Here we demonstrate that, without a calibration step and in the presence of core-to-core coupling, we can obtain fluorescence images with a resolution better than the core-to-core spacing. This is accomplished by taking advantage of the memory effect present in these kinds of fibers.

Enhanced resolution in a multimode fiber imaging system

Multimode fibers have recently been demonstrated to be a promising candidate for ultrathin and high resolution endoscopy. However, this method does not offer depth discrimination for fluorescence imaging and the numerical aperture of the fiber limits its resolution. In this paper we demonstrate optical sectioning and enhanced resolution using saturated excitation and temporal modulation. Using a continuous wave laser excitation, we demonstrate improved resolution in all three dimensions and increased image contrast by rejecting out of focus light.

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.

Optical-resolution photoacoustic imaging through thick tissue with a thin capillary as a dual optical-in acoustic-out waveguide

We demonstrate the ability to guide high-frequency photoacoustic waves through thick tissue with a water-filled silica-capillary (150 μm inner diameter and 30 mm long). An optical-resolution photoacoustic image of a 30 μm diameter absorbing nylon thread was obtained by guiding the acoustic waves in the capillary through a 3 cm thick fat layer. The transmission loss through the capillary was about −20 dB, much lower than the −120 dB acoustic attenuation through the fat layer. The overwhelming acoustic attenuation of high-frequency acoustic waves by biological tissue can therefore be avoided by the use of a small footprint capillary acoustic waveguide for remote detection. We finally demonstrate that the capillary can be used as a dual optical-in acoustic-out waveguide, paving the way for the development of minimally invasive optical-resolution photoacoustic endoscopes free of any acoustic or optical elements at their imaging tip.

High power, ultrashort pulse control through a multi-core fiber for ablation

Ultrashort pulse ablation has become a useful tool for micromachining and biomedical surgical applications. Implementation of ultrashort pulse ablation in confined spaces has been limited by endoscopic delivery and focusing of a high peak power pulse. Here we demonstrate ultrashort pulse ablation through a thin multi-core fiber (MCF) using wavefront shaping, which allows for focusing and scanning the pulse without requiring distal end optics and enables a smaller ablation tool. The intensity necessary for ablation is significantly higher than for multiphoton imaging. We show that the ultimate limitations of the MCF based ablation are the nonlinear effects induced by the pulse in the MCFs cores. We characterize and compare the performance of two devices utilizing a different number of cores and demonstrate ultrashort pulse ablation on a thin film of gold.

Three-dimensional microfabrication through a multimode optical fiber

3D printing based on additive manufacturing is an advanced manufacturing technique that allows the fabrication of arbitrary macroscopic and microscopic objects. Many 3D printing systems require large optical elements or nozzles in proximity to the built structure. This prevents their use in applications in which there is no direct access to the area where the objects have to be printed. Here, we demonstrate three-dimensional microfabrication based on two-photon polymerization (TPP) through an ultra-thin printing nozzle of 560 µm in diameter. Using wavefront shaping, femtosecond infrared pulses are focused and scanned through a multimode optical fiber (MMF) inside a photoresist that polymerizes via two-photon absorption. We show the construction of arbitrary 3D structures built with voxels of diameters down to 400 nm on the other side of the fiber. To our knowledge, this is the first demonstration of microfabrication through a multimode optical fiber. The proposed printing nozzle can reach and manufacture micro-structures in otherwise inaccessible areas through small apertures. Our work represents a new area which we refer to as endofabrication.

Wavefront shaping for ultrashort pulse delivery through optical fibers for imaging and ablation

We demonstrate high power ultrashort pulse delivery through a commercially available multicore fiber (MCF) and a multimode graded-index fiber (GRINF) for imaging and laser ablation. Lensless focusing and digital scanning of ultrashort pulses through the optical fibers is realized using wavefront shaping. We compare the performance of the two systems in terms of focusing efficiency and peak power delivery. Furthermore, we investigate the limitations that nonlinearities induce when high peak power ultrashort pulses are launched in MCFs and GRIN fibers. Proximally-only controlled two-photon fluorescence imaging and laser ablation are demonstrated through both investigated systems.

Ultrashort pulse laser ablation through a multi-core fiber

Ultrashort pulse laser ablation is a useful tool in material processing and biomedical fields [1]. A key challenge to expanding the applications of laser ablation is delivering high peak power, ultrashort laser pulses beyond the reach of conventional microscope objectives. Optical fiber endoscopes are widely used to guide light in confined spaces. However, the endoscopic delivery of a focused ultrashort pulse with peak power high enough to perform laser ablation is susceptible to nonlinear distortions or damage of the fiber itself. Focusing and scanning the pulse through single-mode fibers requires the use of distal optics, which will significantly increase the size of the endoscope. To bypass the current limitations in delivering, focusing and scanning a high peak power ultrashort pulse, we combine multicore fibers (MCFs) with wavefront shaping techniques. In this way, the pulse energy is spread between the thousands of cores of the MCF, thereby mitigating fiber damage and the nonlinearities when delivering an ultrashort pulse of high peak power. Additionally, the use of wavefront shaping techniques allows light to be focused and scanned at the distal end of a fiber without using any distal end optical components, thus maintaining an ultrathin size of the endoscope.