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Hervé Rigneault

Publications and source records attributed to Hervé Rigneault.

At least 37 records · Page 2Linked to original sources

Noise in stimulated Raman scattering measurement: From basics to practice

We revisit laser intensity noise in the context of stimulated Raman scattering (SRS), which has recently proved to be a key technique to provide label free images of chemical bonds in biological and medical samples. Contrary to most microscopy techniques, which detect a weak photon flux resulting from light matter interactions, SRS is a pump-probe scheme that works in the high flux regime and happens as a weak modulation ($10^{-4}-10^{-6}$) in a strong laser field. As a result, laser noise is a key issue in SRS detection. This practical tutorial provides the experimentalists with the tools required to assess the amount of noise and the ultimate SRS detection limit in a conventional lock-in-based SRS system. We first define the quantities that are relevant when discussing intensity noise, and illustrate them through a conventional model of light detection by a photodiode. Stimulated Raman Scattering is then introduced in its lock-in-based implementation, and the model presented is adapted in this particular case. The power spectral density (PSD), relative intensity noise (RIN), signal to noise ratio (SNR), and sensitivity of the system are derived and discussed. Two complementary methods are presented that allow measurement of the RIN and assessment of the performance of a SRS system. Such measurements are illustrated on two commercial laser systems. Finally, the consequences of noise in SRS are discussed, and future developments are suggested. The presentation is made simple enough for under-graduated, graduated students, and newcomers in the field of stimulated Raman, and more generally in pump-probe based schemes.

physics.ins-det↗

Shot noise limited high speed stimulated Raman microscopy

We report a shot noise limited high speed stimulated Raman microscopy platform allowing to acquire molecular vibrational spectra over \SI{200}{\per \cm} in \SI{12}{\micro \second} at a scan rate of 40kHz. Using spectral focusing together with optimized acousto-optics programmable dispersive filters the designed low noise imaging platform performs chemical imaging of dynamical processes such as Mannitol crystal hydratation and reaches a signal to noise ratio sufficient to perform label free histological imaging on frozen human colon tissue slides.

physics.optics↗

Flexible lensless endoscope with a conformationally invariant multi-core fiber

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 letter we report for the first time 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.

physics.app-ph↗

Fast stimulated Raman imaging for intraoperative gastro-intestinal cancer detection

Conventional haematoxylin, eosin and saffron (HES) histopathology, currently the gold-standard for pathological diagnosis of cancer, requires extensive sample preparations that are achieved within time scales that are not compatible with intra-operative situations where quick decisions must be taken. Providing to pathologists a close to real-time technology revealing tissue structures at the cellular level with HES histologic quality would provide an invaluable tool for surgery guidance with evident clinical benefit. Here, we specifically develop a stimulated Raman imaging based framework that demonstrates gastro-intestinal (GI) cancer detection of unprocessed human surgical specimens. The generated stimulated Raman histology (SRH) images combine chemical and collagen information to mimic conventional HES histopathology staining. We report excellent agreements between SRH and HES images acquire on the same patients for healthy, pre-cancerous and cancerous colon and pancreas tissue sections. We also develop a novel fast SRH imaging modality that captures at the pixel level all the information necessary to provide instantaneous SRH images. These developments pave the way for instantaneous label free GI histology in an intra-operative context.

physics.med-ph↗

Arago spot formation in the time domain

Using the space-time duality we theoretically and experimentally revisit the Arago spot formation in the time domain and explore the temporal emergence of a bright spot in the time shadow of an initial waveform. We describe the linear regime of propagation using Fresnel's integrals and show that in the presence of Kerr nonlinearity the Arago spot formation is affected depending on the sign of the dispersion. We finally confirm experimentally the Arago spot formation in the time domain using a telecommunication optical fiber platform.

physics.optics↗

Compressive Sampling Approach for Image Acquisition with Lensless Endoscope

The lensless endoscope is a promising device designed to image tissues in vivo at the cellular scale. The traditional acquisition setup consists in raster scanning during which the focused light beam from the optical fiber illuminates sequentially each pixel of the field of view (FOV). The calibration step to focus the beam and the sampling scheme both take time. In this preliminary work, we propose a scanning method based on compressive sampling theory. The method does not rely on a focused beam but rather on the random illumination patterns generated by the single-mode fibers. Experiments are performed on synthetic data for different compression rates (from 10 to 100% of the FOV).

cs.CV↗

Optical Antenna-based Fluorescence Correlation Spectroscopy to Probe the Nanoscale Dynamics of Biological Membranes

The plasma membrane of living cells is compartmentalized at multiple spatial scales ranging from the nano- to the meso-scale. This non-random organization is crucial for a large number of cellular functions. At the nanoscale, cell membranes organize into dynamic nano-assemblies enriched by cholesterol, sphingolipids and certain types of proteins. Investigating these nano-assemblies known as lipid rafts is of paramount interest in fundamental cell biology. However, this goal requires simultaneous nanometer spatial precision and microsecond temporal resolution which is beyond the reach of common microscopes. Optical antennas based on metallic nanostructures efficiently enhance and confine light into nanometer dimensions, breaching the diffraction limit of light. In this Perspective, we discuss recent progress combining optical antennas with fluorescence correlation spectroscopy (FCS) to monitor microsecond dynamics at nanoscale spatial dimensions. These new developments offer numerous opportunities to investigate lipid and protein dynamics in both mimetic and native biological membranes.

physics.bio-ph↗

Transient Nanoscopic Phase Separation in Biological Lipid Membranes Resolved by Planar Plasmonic Antennas

Nanoscale membrane assemblies of sphingolipids, cholesterol, and certain proteins, also known as lipid rafts, play a crucial role in facilitating a broad range of important cell functions. Whereas on living cell membranes lipid rafts have been postulated to have nanoscopic dimensions and to be highly transient, the existence of a similar type of dynamic nanodomains in multicomponent lipid bilayers has been questioned. Here, we perform fluorescence correlation spectroscopy on planar plasmonic antenna arrays with different nanogap sizes to assess the dynamic nanoscale organization of mimetic biological membranes. Our approach takes advantage of the highly enhanced and confined excitation light provided by the nanoantennas together with their outstanding planarity to investigate membrane regions as small as 10 nm in size with microsecond time resolution. Our diffusion data are consistent with the coexistence of transient nanoscopic domains in both the liquid-ordered and the liquid-disordered microscopic phases of multicomponent lipid bilayers. These nanodomains have characteristic residence times between 30 and 150 μs and sizes around 10 nm, as inferred from the diffusion data. Thus, although microscale phase separation occurs on mimetic membranes, nanoscopic domains also coexist, suggesting that these transient assemblies might be similar to those occurring in living cells, which in the absence of raft-stabilizing proteins are poised to be short-lived. Importantly, our work underscores the high potential of photonic nanoantennas to interrogate the nanoscale heterogeneity of native biological membranes with ultrahigh spatiotemporal resolution.

physics.bio-ph↗

Planar Optical Nanoantennas Resolve Cholesterol-Dependent Nanoscale Heterogeneities in the Plasma Membrane of Living Cells

Optical nanoantennas can efficiently confine light into nanoscopic hotspots, enabling single-molecule detection sensitivity at biological relevant conditions. This innovative approach to breach the diffraction limit offers a versatile platform to investigate the dynamics of individual biomolecules in living cell membranes and their partitioning into cholesterol-dependent lipid nanodomains. Here, we present optical nanoantenna arrays with accessible surface hotspots to study the characteristic diffusion dynamics of phosphoethanolamine (PE) and sphingomyelin (SM) in the plasma membrane of living cells at the nanoscale. Fluorescence burst analysis and fluorescence correlation spectroscopy performed on nanoantennas of different gap sizes show that, unlike PE, SM is transiently trapped in cholesterol-enriched nanodomains of 10 nm diameter with short characteristic times around 100 μs. The removal of cholesterol led to the free diffusion of SM, consistent with the dispersion of nanodomains. Our results are consistent with the existence of highly transient and fluctuating nanoscale assemblies enriched by cholesterol and sphingolipids in living cell membranes, also known as lipid rafts. Quantitative data on sphingolipids partitioning into lipid rafts is crucial to understand the spatiotemporal heterogeneous organization of transient molecular complexes on the membrane of living cells at the nanoscale. The proposed technique is fully biocompatible and thus provides various opportunities for biophysics and live cell research to reveal details that remain hidden in confocal diffraction-limited measurements.

physics.bio-ph↗

Wavefront-sensing with a thin diffuser

We propose and implement a broadband, compact, and low-cost wavefront sensing scheme by simply placing a thin diffuser in the close vicinity of a camera. The local wavefront gradient is determined from the local translation of the speckle pattern. The translation vector map is computed thanks to a fast diffeomorphic image registration algorithm and integrated to reconstruct the wavefront profile. The simple translation of speckle grains under local wavefront tip/tilt is ensured by the so-called "memory effect" of the diffuser. Quantitative wavefront measurements are experimentally demonstrated both for the few first Zernike polynomials and for phase-imaging applications requiring high resolution. We finally provided a theoretical description of the resolution limit that is supported experimentally.

physics.optics↗

Origin and suppression of parasitic signals in Kagomé lattice hollow core fibers used for SRS microscopy and endoscopy

Hollow core fibers are considered as promising candidates to deliver intense temporally overlapping picosecond pulses in applications such as stimulated Raman scattering (SRS) microscopy and endoscopy because of their inherent low nonlinearity compared to solid-core silica fibers. Here we demonstrate that, contrary to prior assumptions, parasitic signals are generated in Kagomé lattice hollow core fibers. We identify the origin of the parasitic signals as an interplay between the Kerr nonlinearity of air and frequency-dependent fiber losses. Importantly, we identify the special cases of experimental parameters that are free from parasitic signals, making hollow core fibers ideal candidates for noise-free SRS microscopy and endoscopy.

physics.optics↗

High-resolution multimodal flexible coherent Raman endoscope

Coherent Raman scattering microscopy is a fast, label-free and chemically specific imaging technique that has a high potential for future in-vivo optical histology. However, its imaging depth into tissues is limited to the sub-millimeter range by absorption and scattering. Performing coherent Raman imaging in a fiber endoscope system is a crucial step to image deep inside living tissues and provide the information inaccessible with current microscopy tools. However the development of coherent Raman endoscopy has been hampered by several issues in the fiber delivery of the excitation pulses and signal collection. Here, we present a flexible, compact, 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 allows distortion-less, background-free delivery of femtosecond 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 with 0.8 μm resolution over a field of view up to 320 μm and at a rate of 0.8 frames/s. These results pave the way for intra-operative label-free imaging applied to real-time histopathology diagnosis and surgery guidance.

physics.med-ph↗

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.

physics.optics↗

Complementary Speckle Patterns : deterministic interchange of intrinsic vortices and maxima through Scattering Media

Intensity minima and maxima of speckle patterns obtained behind a diffuser are experimentally interchanged by applying a spiral phase delay of charge $\pm 1$ to the impinging coherent beam. This transform arises from the intuitive expectation that a tightly focused beam is so-changed into a vortex beam and vice-versa. The statistics of extrema locations and the intensity distribution of the so-generated "complementary" patterns are characterized by numerical simulations. It is demonstrated experimentally that the incoherent superposition of the three "complementary speckle patterns" yield a synthetic speckle grain size enlarged by a factor $\sqrt{3}$. A cyclic permutation of optical vortices and maxima is unexpectedly observed and discussed.

physics.optics↗

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 MCF experimental feasibility. We demonstrate experimentally pixelation-free lensless endoscopy imaging over a 120 micron 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.

physics.optics↗

Widefield lensless endoscopy via speckle-correlations

Flexible fiber-optic endoscopes provide a minimally-invasive solution for imaging at depths beyond the reach of conventional microscopes. Current endoscopes require focusing and/or scanning mechanisms at the distal end, which limit miniaturization and frame-rate, and induce aberrations. Alternative lensless solutions are based on adaptive wavefront-correction, but are extremely sensitive to fiber bending. Here, we demonstrate a novel endoscopic approach, which enables single-shot imaging at a variable working distance through a conventional fiber bundle, without the use of any distal optics. Our approach computationally retrieves the object image from a single speckle pattern transmitted through the bundle, exploiting phase information preserved through inherent angular speckle correlations. Unlike conventional fiber-bundle endoscopes, the resulting image is unpixelated, the resolution is diffraction-limited, objects can be imaged at variable working distance, and the technique is completely insensitive to fiber bending. Since no optical elements are required, miniaturization is limited only by the bundle diameter.

physics.optics↗