Subcellular spatial resolution achieved for deep-brain imaging in vivo using a minimally invasive multimode fiber

Achieving intravital optical imaging with diffraction-limited spatial resolution of deep-brain structures represents an important step toward the goal of understanding the mammalian central nervous system1,2,3,4. Advances in wavefront-shaping methods and computational power have recently allowed for a novel approach to high-resolution imaging, utilizing deterministic light propagation through optically complex media and, of particular importance for this work, multimode optical fibers (MMFs)5,6,7. We report a compact and highly optimized approach for minimally invasive in vivo brain imaging applications. The volume of tissue lesion was reduced by more than 100-fold, while preserving diffraction-limited imaging performance utilizing wavefront control of light propagation through a single 50-μm-core MMF. Here, we demonstrated high-resolution fluorescence imaging of subcellular neuronal structures, dendrites and synaptic specializations, in deep-brain regions of living mice, as well as monitored stimulus-driven functional Ca2+ responses. These results represent a major breakthrough in the compromise between high-resolution imaging and tissue damage, heralding new possibilities for deep-brain imaging in vivo

Three-dimensional holographic optical manipulation through a high-numerical-aperture soft-glass multimode fibre

Holographic optical tweezers (HOT) hold great promise for many applications in biophotonics, allowing the creation and measurement of minuscule forces on biomolecules, molecular motors and cells. Geometries used in HOT currently rely on bulk optics, and their exploitation in vivo is compromised by the optically turbid nature of tissues. We present an alternative HOT approach in which multiple three-dimensional (3D) traps are introduced through a high-numerical-aperture multimode optical fibre, thus enabling an equally versatile means of manipulation through channels having cross-section comparable to the size of a single cell. Our work demonstrates real-time manipulation of 3D arrangements of micro-objects, as well as manipulation inside otherwise inaccessible cavities. We show that the traps can be formed over fibre lengths exceeding 100 mm and positioned with nanometric resolution. The results provide the basis for holographic manipulation and other high-numerical-aperture techniques, including advanced microscopy, through single-core-fibre endoscopes deep inside living tissues and other complex environments

GPU accelerated holography for multimode fiber applications

We demonstrate a compact GPU accelerated digital holography system for beam-shaping applications at the end of multimode fibre. The complete control of phase and amplitude on phase-only SLM is achieved through GPU implemented Gerchberg-Saxton and Yang-Gu algorithms. The GPU implementation is two orders of magnitude faster than the CPU implementation which allows video-rate image control at the distal end of the fiber virtually free of interference effects. We discuss the implementation of the algorithms and possible applications of the technology for structured illumination imaging at the end of multimode fibre.6 page(s

Compact multimode fiber beam-shaping system based on GPU accelerated digital holography

Real-time, on-demand, beam shaping at the end of the multimode fiber has recently been made possible by exploiting the computational power of rapidly evolving graphics processing unit (GPU) technology [Opt. Express 22, 2933 (2014)]. However, the current state-of-the-art system requires the presence of an acousto-optic deflector (AOD) to produce images at the end of the fiber without interference effects between neighboring output points. Here, we present a system free from the AOD complexity where we achieve the removal of the undesired interference effects computationally using GPU implemented Gerchberg-Saxton and Yang-Gu algorithms. The GPU implementation is two orders of magnitude faster than the CPU implementation which allows video-rate image control at the distal end of the fiber virtually free of interference effects.4 page(s

Multimode fibres:a pathway towards deep-tissue fluorescence microscopy

Fluorescence microscopy has emerged as a pivotal platform for imaging in the life sciences. In recent years, the overwhelming success of its different modalities has been accompanied by various efforts to carry out imaging deeper inside living tissues. A key challenge of these efforts is to overcome scattering and absorption of light in such environments. Multiple strategies (e.g. multi-photon, wavefront correction techniques) extended the penetration depth to the current state-of-the-art of about 1000μm at the resolution of approximately 1μm. The only viable strategy for imaging deeper than this is by employing a fibre bundle based endoscope. However, such devices lack resolution and have a significant footprint (1mm in diameter), which prohibits their use in studies involving tissues deep in live animals. We have recently demonstrated a radically new approach that delivers the light in/out of place of interest through an extremely thin (tens of microns in diameter) cylindrical glass tube called a multimode optical fibre (MMF). Not only is this type of delivery much less invasive compared to fibre bundle technology, it also enables higher resolution and has the ability to image at any plane behind the fibre without any auxiliary optics. The two most important limitations of this exciting technology are (i) the lack of bending flexibility and (ii) high demands on computational power, making the performance of such systems slow. We will discuss how to overcome these limitations.6 page(s

Untangled modes in multimode waveguides

Small, fibre-based endoscopes have already improved our ability to image deep within the human body. A novel approach introduced recently utilised disordered light within a standard multimode optical fibre for lensless imaging. Importantly, this approach brought very significant reduction of the instruments footprint to dimensions below 100 μm. The most important limitations of this exciting technology is the lack of bending flexibility - imaging is only possible as long as the fibre remains stationary. The only route to allow flexibility of such endoscopes is in trading-in all the knowledge about the optical system we have, particularly the cylindrical symmetry of refractive index distribution. In perfect straight step-index cylindrical waveguides we can find optical modes that do not change their spatial distribution as they propagate through. In this paper we present a theoretical background that provides description of such modes in more realistic model of real-life step-index multimode fibre taking into account common deviations in distribution of the refractive index from its ideal step-index profile. Separately, we discuss how to include the influence of fibre bending

Media 2: Compact multimode fiber beam-shaping system based on GPU accelerated digital holography

Originally published in Optics Letters on 15 January 2015 (ol-40-2-197

Untangled modes in multimode fibres for flexible microendoscopy

Small, fibre-based endoscopes have already improved our ability to image deep within the human body. A novel approach introduced recently utilised disordered light within a standard multimode optical fibre for lensless imaging. Importantly, this approach brought very significant reduction of the instruments footprint to dimensions below 100 μm. The most important limitations of this exciting technology is the lack of bending flexibility - imaging is only possible as long as the fibre remains stationary. The only route to allow flexibility of such endoscopes is in trading-in all the knowledge about the optical system we have, particularly the cylindrical symmetry. In perfect cylindrical waveguides we can find optical modes that do not change their spatial distribution as they propagate through. We show that typical fibers retain such highly ordered propagation of light over remarkably large distances, which allows correction operators to be introduced in imaging geometries in order to maintain high-quality performance even in such flexible micro-endoscopes.8 page(s

Media 1: Compact multimode fiber beam-shaping system based on GPU accelerated digital holography

Originally published in Optics Letters on 15 January 2015 (ol-40-2-197

Fibre-based imaging:new challenges

A novel minimally invasive micro-endoscopes utilizing disordered light within a standard multimode optical fibre have been introduced recently. The two most important limitations of this exciting technology are (i) the lack of bending flexibility (transformation matrix is only valid as long as the fibre remains stationary) and (ii) high demands on computational power, making the performance of such systems slow. Here we discuss possible routes to address the later one: We introduce a GPU toolbox to make this powerful technique faster and more accessible to bio-medical researchers.Publisher PD