Non-invasive optical focusing inside scattering media is still a big challenge because inhomogeneous media scatter both incoming photons for focusing and outgoing photons for observation. Various approaches, utilizing non-linear fluorescence or ultrasound, have been reported to address this difficulty. However, implementation of these methods is complicated and highly expensive, as ultrafast laser systems or photo-acoustic equipment must be employed. Here, we demonstrate a wavefront shaping technique to achieve non-invasive focusing (NiF) inside scattering media using only a linear fluorescent signal. Contrast and mean of incoherent speckles, produced by the linear fluorescence, are utilized as feedback signals to optimize the input wavefront. While increasing speckle contrast makes the focus tighter, and increasing the speckle mean enhances the intensity, fine-tuning the contribution of these two factors in our two-step optimization is essential. An optimal wavefront is found to achieve simultaneously both a micrometer focal spot size (down to 20 um diameter) and high intensity (more than a 100-fold enhancement) inside the scattering media. Our method promises a new route in life science towards focusing, imaging or manipulating deep into biological tissues with linear fluorescent agents.
Fluorescence is a powerful mean to probe information processing in the mammalian brain. However, neuronal tissues are highly heterogeneous and thus opaque to light. A wide set of non-invasive or invasive techniques for scattered light rejection, optical sectioning or localized excitation, have been developed, but non-invasive optical recording of activity through highly scattering layer beyond the ballistic regime is to date impossible. Here, we show that functional signals from fluorescent time-varying sources located below an highly scattering tissue can be retrieved efficiently, by exploiti…
In biological microscopy, light scattering represents the main limitation to image at depth. Recently, a set of wavefront shaping techniques has been developed in order to manipulate coherent light in strongly disordered materials. The Transmission Matrix approach has shown its capability to inverse the effect of scattering and efficiently focus light. In practice, the matrix is usually measured using an invasive detector or low-resolution acoustic guide stars. Here, we introduce a non-invasive and all-optical strategy based on linear fluorescence to reconstruct the transmission matrices, to a…
A major open challenge in neuroscience is the ability to measure and perturb neural activity in vivo from well-defined neural sub-populations at cellular resolution anywhere in the brain. However, limitations posed by scattering and absorption prohibit non-invasive (surface) multiphoton approaches for deep (>2mm) structures, while Gradient Refreactive Index (GRIN) endoscopes are thick and cause significant damage upon insertion. Here, we demonstrate a novel microendoscope to image neural activity at arbitrary depths via an ultrathin multimode optical fiber (MMF) probe that is 5-10X thinner tha…
Extending super-resolution imaging techniques to objects hidden in strongly scattering media potentially revolutionize the technical analysis for much broader categories of samples, such as biological tissues. The main challenge is the media's inhomogeneous structures which scramble the light path and create noise-like speckle patterns, hindering the object's visualization even at a low-resolution level. Here, we propose a computational method relying on the object's spatial and temporal fluctuation to visualize nanoscale objects through scattering media non-invasively. The fluctuating object …
Computational Cannula Microscopy (CCM) is a high-resolution widefield fluorescence imaging approach deep inside tissue, which is minimally invasive. Rather than using conventional lenses, a surgical cannula acts as a lightpipe for both excitation and fluorescence emission, where computational methods are used for image visualization. Here, we enhance CCM with artificial neural networks to enable 3D imaging of cultured neurons and fluorescent beads, the latter inside a volumetric phantom. We experimentally demonstrate transverse resolution of ~6um, field of view ~200um and axial sectioning of ~…
Optical imaging deep inside scattering media remains a fundamental problem in bio-imaging. While wavefront shaping has been shown to allow focusing of coherent light at depth, achieving it non-invasively remains a challenge. Various feedback mechanisms, in particular acoustic or non-linear fluorescence-based, have been put forward for this purpose. Non-invasive focusing at depth on fluorescent objects with linear excitation is, however, still unresolved. Here we report a simple method for focusing inside a scattering medium in an epi-detection geometry with a linear signal: optimizing the spat…
We present a method for single-shot three-dimensional imaging through scattering media with a three-dimensional memory effect. In the proposed computational process, a captured speckle image is two-dimensionally correlated with different scales, and the object is three-dimensionally recovered with three-dimensional phase retrieval. Our method was experimentally demonstrated with a lensless setup and was compared with a multi-shot approach used in our previous work [Y. Okamoto, et al., Opt. Lett. 44, 2526-2529 (2019)].
Super-resolution imaging with advanced optical systems has been revolutionizing technical analysis in various fields from biological to physical sciences. However, many objects are hidden by strongly scattering media such as rough wall corners or biological tissues that scramble light paths, create speckle patterns and hinder object's visualization, let alone super-resolution imaging. Here, we realize a method to do non-invasive super-resolution imaging through scattering media based on stochastic optical scattering localization imaging (SOSLI) technique. Simply by capturing multiple speckle p…
Optical imaging through complex media, such as biological tissues or fog, is challenging due to light scattering. In the multiple scattering regime, wavefront shaping provides an effective method to retrieve information; it relies on measuring how the propagation of different optical wavefronts are impacted by scattering. Based on this principle, several wavefront shaping techniques were successfully developed, but most of them are highly invasive and limited to proof-of-principle experiments. Here, we propose to use a neural network approach to non-invasively characterize and control light sc…
We develop a method based on the cross-spectrum of an intensity-modulated CW laser, which can extract a signal from an extremely noisy environment and image objects hidden in turbid media. We theoretically analyzed our scheme and performed the experiment by scanning the object placed in between two ground glass diffusers. The image of the object is retrieved by collecting the amplitudes at the modulation frequency of all the cross-spectra. Our method is non-invasive, easy-to-implement, and can work for both static and dynamic media.
We demonstrate focusing and imaging through a scattering medium noninvasively by using wavefront shaping. Our concept is based on utilizing the spatial fluorescence contrast which naturally exists in the hidden target object. By scanning the angle of incidence of the illuminating laser beam and maximizing the variation of the detected fluorescence signal from the object, as measured by a bucket detector at the front of the scattering medium, we are able to generate a tightly focused excitation spot. Thereafter, an image is obtained by scanning the focus over the object. The requirements for ap…
Several matrix approaches were developed to control light propagation through multiple scattering media under illumination of ultrashort pulses of light. These matrices can be recorded either with spectral or temporal resolution. Thanks to wavefront shaping, temporal and spatial refocusing have been demonstrated. In this work, we study how these different methods can be exploited to enhance a two-photon excitation fluorescence process. We first compare the different techniques on micrometer-size isolated fluorescent beads. We then demonstrate point-scanning imaging of such fluorescent microbea…
On-invasive optical imaging techniques are essential diagnostic tools in many fields. Although various recent methods have been proposed to utilize and control light in multiple scattering media, non-invasive optical imaging through and inside scattering layers across a large field of view remains elusive due to the physical limits set by the optical memory effect, especially without wavefront shaping techniques. Here, we demonstrate an approach that enables non-invasive fluorescence imaging behind scattering layers with field-of-views extending well beyond the optical memory effect. The metho…
Optical imaging through scattering media is a long-standing challenge. Although many approaches have been developed to focus light or image objects through scattering media, they are either invasive, restricted to stationary or slowly-moving media, or require high-resolution cameras and complex algorithms to retrieve the images. Here we introduce a computational imaging technique that can overcome these restrictions by exploiting spatial-temporal encoded patterns (STEP). We present non-invasive imaging through scattering media with a single-pixel photodetector. We show that the method is insen…
A holistic approach using non-invasive multimodal imaging and spectroscopic techniques to study the materials (pigments, drawing materials and paper) and painting techniques of watercolour paintings is presented. The non-invasive imaging and spectroscopic techniques include VIS-NIR reflectance spectroscopy and multispectral imaging, micro-Raman spectroscopy, X-ray fluorescence spectroscopy (XRF) and optical coherence tomography (OCT). The three spectroscopic techniques complement each other in pigment identification. Multispectral imaging (near infrared bands), OCT and micro-Raman complement e…
When waves propagate through a strongly scattering medium the energy is transferred to the incoherent wave part by scattering. The wave intensity then forms a random speckle pattern seemingly without much useful information. However, a number of recent physical experiments show how one can extract useful information from this speckle pattern. Here we present the mathematical analysis that explains the quite stunning performance of such a scheme for speckle imaging. Our analysis identifies a scaling regime where the scheme works well. This regime is the white-noise paraxial regime, which leads …
EvidenceMoE: A Physics-Guided Mixture-of-Experts with Evidential Critics for Advancing Fluorescence Light Detection and Ranging in Scattering Media
Center for Modeling, Simulation, & Imaging in Medicine（建模、仿真与成像医学中心）；Rensselaer Polytechnic Institute（伦斯勒理工学院）；University at Albany（阿尔巴尼大学）；IBM T.J. Watson Research Center（IBM 沃森研究中心）
Laser Pulse Diagnostics of Ultrafast <8 fs Pulses Through Two-Photon Absorption Fluorescence In Liquid Media -- The Role of GVD and Third-Order Dispersion
$μ$NeuFMT: Optical-Property-Adaptive Fluorescence Molecular Tomography via Implicit Neural Representation
School of Information Science and Technology, ShanghaiTech University（信息科学与技术学院，上海科技大学）；School of Mathematics, University of Birmingham（数学学院，伯明翰大学）；School of Physical Science and Technology, ShanghaiTech University（物理科学与技术学院，上海科技大学）；Institute of Medical Robotics, School of Biomedical Engineering, Shanghai Jiao Tong University（医学机器人研究所，生物医学工程学院，上海交通大学）；State Key Laboratory of Advanced Medical Materials and Devices, ShanghaiTech University（先进医学材料与器件国家重点实验室，上海科技大学）
We propose a new method to image fluorescent objects through turbid media base on Airy beam scanning. This is achieved by using the non-diffractive nature of Airy beams, namely their ability to maintain their shape while penetrating through a highly scattering medium. We show, that our technique can image fluorescent objects immersed in turbid media with higher resolution and signal to noise than confocal imaging. As proof-of-principle, we demonstrate imaging of 1$μ$m sized fluorescent beads through a dense suspension of yeast cells with an attenuation coefficient of 51cm$^{-1}$ at a depth of …
The efficient optical tool for elimination of the phase and amplitude distortions produced by imperfectness of the optical elements in microscope lightpath considered. This robust procedure described by simple theoretical model proved to be successful to repair the noisy images with noise to signal ratio close to 100. It is shown that Van Cittert - Zernike theorem provides adequate description of imperfect microscope operating under both coherent and incoherent illumination conditions and having turbid media in object and intermediate planes.
The ability to steer light propagation inside scattering media has long been sought-after due to its potential widespread applications. To form optical foci inside scattering media, the only feasible strategy is to guide photons by using either implanted or virtual guide stars. However, all of these guide stars must be introduced extrinsically, either invasively or by physical contact, limiting the scope of their application. Here, we focus light inside scattering media by employing intrinsic dynamics as guide stars. By time-reversing the perturbed component of the scattered light adaptively, …
Scattering can rapidly degrade our ability to form an optical image, to the point where only speckle-like patterns can be measured. Truly non-invasive imaging through a strongly scattering obstacle is difficult, and usually reliant on a computationally intensive numerical reconstruction. In this work we show that, by combining the cross-correlations of the measured speckle pattern at different times, it is possible to track a moving object with minimal computational effort and over a large field of view.
Optical imaging through complex scattering media is one of the major technical challenges with important applications in many research fields, ranging from biomedical imaging, astronomical telescopy, and spatially multiplex optical communications. Although various approaches for imaging though turbid layer have been recently proposed, they had been limited to two-dimensional imaging. Here we propose and experimentally demonstrate an approach for three-dimensional single-shot imaging of objects hidden behind an opaque scattering layer. We demonstrate that under suitable conditions, it is possib…
In turbid media, scattering of light scrambles information of the incident beam and represents an obstacle to optical imaging. Noninvasive imaging through opaque layers is challenging for dynamic and wide-field objects due to unreliable image reconstruction processes. We here propose a new perspective to solve these problems: rather than using the full point-spread-function (PSF), the wave distortions in scattering layers can be characterized with only the phase of the optical-transfer-function (OTF, the Fourier transform of PSF), with which diffraction-limit images can be analytically solved.…
Fluorescence microscopy is widely used in biological imaging, however scattering from tissues strongly limits its applicability to a shallow depth. In this work we adapt a methodology inspired from stellar speckle interferometry, and exploit the optical memory effect to enable fluorescence microscopy through a turbid layer. We demonstrate efficient reconstruction of micrometer-size fluorescent objects behind a scattering medium in epi-microscopy, and study the specificities of this imaging modality (magnification, field of view, resolution) as compared to traditional microscopy. Using a modifi…
We present a minimally-invasive endoscope based on a multimode fiber that combines photoacoustic and fluorescence sensing. From the measurement of a transmission matrix during a prior calibration step, a focused spot is produced and raster-scanned over a sample at the distal tip of the fiber by use of a fast spatial light modulator. An ultra-sensitive fiber-optic ultrasound sensor for photoacoustic detection placed next to the fiber is combined with a photodetector to obtain both fluorescence and photoacoustic images with a distal imaging tip no larger than 250um. The high signal-to-noise rati…
Optical wavefront-shaping has emerged as a powerful tool to manipulate light in strongly scattering media. It enables diffraction-limited focusing and imaging at depths where conventional microscopy techniques fail. However, while most wavefront-shaping works to-date exploited direct access to the target or implanted probes, the challenge is to apply it non-invasively inside complex samples. Ultrasonic-tagging techniques have been recently demonstrated but these require a sequential point-by- point acquisition, a major drawback for imaging applications. Here, we introduce a novel approach to n…
Fluorescence microscopy has revolutionized biomedical research over the past three decades. Its high molecular specificity and unrivaled single molecule level sensitivity have enabled breakthroughs in a variety of research fields. For in vivo applications, its major limitation is the superficial imaging depth as random scattering in biological tissues causes exponential attenuation of the ballistic component of a light wave. Here we present fluorescence microscopy beyond the ballistic regime by combining single cycle pulsed ultrasound modulation and digital optical phase conjugation. We demons…
Non-degradable fluorophores that accumulate as Lipofuscin in retinal pigment epithelium (RPE) cells has been a major source of intrinsic biomarker for quantifying the progression of several diseases, including age-related macular degeneration, Stargardt disease and, more. Recent progression in quantifying these diseases is entertained mostly by a few noninvasive imaging techniques, relied either on the life-time of the retinoid-fluorophores or, their linear and, non-linear absorption cross-sections. Apart from these intrinsic properties, a native, excited state dipole-dipole interaction mediat…
Focusing light into opaque random or scattering media such as biological tissue is a much sought-after goal for biomedical applications such as photodynamic therapy, optical manipulation, and photostimulation. However, focusing with conventional lenses is restricted to one transport mean free path in scattering media, limiting both optical penetration depth and resolution. Focusing deeper is possible by using optical phase conjugation or wavefront shaping to compensate for the scattering. For practical applications, wavefront shaping offers the advantage of a robust optical system that is less…
The much sought after optical transistor --the photonic counterpart of the electronic transistor-- is poised to become a central ingredient in the development of optical signal processing. The motivation for using photons rather than electrons not only comes from their faster dynamics but also from their lower crosstalk and minor environmental decoherence, which enable a high degree of integration and the realization of quantum operations. A single-molecule transistor has been recently demonstrated at cryogenic temperatures. Here, we demonstrate that a single NV centre at room temperature can …
We experimentally demonstrate optical control of filamentation that occurs during propagation of intense, ultrashort laser pulses through crystals like barium fluoride and sapphire. Control is exercised by rotating the plane of polarization of the incident laser radiation and is demonstrated by directly visualizing filamentation in the bulk via six-photon absorption-induced fluorescence and, concomitantly, by probing the spectral and spatial properties of white light that is generated.
Imaging with optical resolution through and inside complex samples is a difficult challenge with important applications in many fields. The fundamental problem is that inhomogeneous samples, such as biological tissues, randomly scatter and diffuse light, impeding conventional image formation. Despite many advancements, no current method enables to noninvasively image in real-time using diffused light. Here, we show that owing to the memory-effect for speckle correlations, a single image of the scattered light, captured with a standard high-resolution camera, encodes all the information that is…
We introduce a generalized version of phase retrieval called multiplexed phase retrieval. We want to recover the phase of amplitude-only measurements from linear combinations of them. This corresponds to the case in which multiple incoherent sources are sampled jointly, and one would like to recover their individual contributions. We show that a recent spectral method developed for phase retrieval can be generalized to this setting, and that its performance follows a phase transition behavior. We apply this new technique to light focusing at depth in a complex medium. Experimentally, although …
Focusing scattered light using wavefront shaping provides interesting perspectives to image deep in opaque samples, as e.g. in nonlinear fluorescence microscopy. Applying these technics to in vivo imaging remains challenging due to the short decorrelation time of the speckle in depth, as focusing and imaging has to be achieved within the order of the decorrelation time. In this paper, we experimentally study the focus lifetime after focusing through dynamical scattering media, when iterative wavefront optimization and speckle decorrelation occur over the same timescale. We show experimental si…
Complete polarimeters deliver the full polarization transfer matrix of a medium that relates input polarization states to output polarization states. In order to interpret the Mueller matrix of a luminescent medium at the emission frequency, accountings are required for polarization transformations of the medium at the excitation frequency, the light scattering event, and the polarization transformations at the emission frequency. A general framework for this kind of analysis is presented herein. The fluorescence Mueller matrix is expressed as a product of Mueller matrices of the medium at the…
Light scattering in inhomogeneous media induces wavefront distortions which pose an inherent limitation in many optical applications. Examples range from microscopy and nanosurgery to astronomy. In recent years, ongoing efforts have made the correction of spatial distortions possible by wavefront shaping techniques. However, when ultrashort pulses are employed scattering induces temporal distortions which hinder their use in nonlinear processes such as in multiphoton microscopy and quantum control experiments. Here we show that correction of both spatial and temporal distortions can be attaine…
We demonstrate the preservation of time-energy entanglement of near-IR photons through thick biological media ($\leq$1.55 mm) and tissue ($\leq$ 235 $μ$m) at room temperature. Using a Franson-type interferometer, we demonstrate interferometric contrast of over 0.9 in skim milk, 2% milk, and chicken tissue. This work supports the many proposed opportunities for nonclassical light in biological imaging and analyses from sub-shot noise measurements to entanglement-enhanced fluorescence imaging, clearly indicating that the entanglement characteristics of photons can be maintained even after propag…
Diffraction-limited imaging through complex scattering media is a long sought after goal with important applications in biomedical research. In recent years, high resolution wavefront-shaping has emerged as a powerful approach to generate a sharp focus through highly scattering, visually opaque samples. However, it requires a localized feedback signal from the target point of interest, which necessitates an invasive procedure in all-optical techniques. Here, we show that by exploiting optical nonlinearities, a diffraction-limited focus can be formed inside or through a complex sample, even whe…
The small correction volume for conventional wavefront shaping methods limits their applications in biological imaging through scattering media. We demonstrate large volume wavefront shaping through a scattering layer with a single correction by conjugate adaptive optics and remote focusing (CAORF). The remote focusing module can keep the conjugation between the AO and scattering layer during three-dimensional scanning. This new configuration provides a wider correction volume by the best utilization of the memory effect in a fast three-dimensional laser scanning microscope. Our results show t…
Non-invasive and single-shot holographic imaging through complex media is technically challenging due to random light scattering which significantly scrambles optical information. Recently, several methods have been presented to address this issue. However, they require complicated measurements of optical transmission matrices, or existing techniques do only retrieve intensity information. Here we propose and experimentally demonstrate a holographic approach for single-shot imaging through a scattering layer based on digital in-line holography in combination with the autocorrelation of the spe…
Optical time-reversal techniques are being actively developed to focus light through or inside opaque scattering media. When applied to biological tissue, these techniques promise to revolutionize biophotonics by enabling deep-tissue non-invasive optical imaging, optogenetics, optical tweezers and photodynamic therapy. In all previous optical time-reversal experiments, the scattered light field was well-sampled during wavefront measurement and wavefront reconstruction, following the Nyquist sampling criterion. Here, we overturn this conventional practice by demonstrating that even when the sca…
We present a stochastic description of multiple scattering of polarized waves in the regime of forward scattering. In this regime, if the source is polarized, polarization survives along a few transport mean free paths, making it possible to measure an outgoing polarization distribution. We solve the direct problem using compound Poisson processes on the rotation group SO(3) and non-commutative harmonic analysis. The obtained solution generalizes previous works in multiple scattering theory and is used to design an algorithm solving the inverse problem of estimating the scattering properties o…
From the earth's crust to the human brain, remitted waves are used for sensing and imaging in a diverse range of diffusive media. Separating the source and detector increases the penetration depth of remitted light, yet rapidly decreases the signal strength, leading to a poor signal-to-noise ratio. Here, we experimentally and numerically show that wavefront shaping a laser beam incident on a diffusive sample enables an order of magnitude remission enhancement, with a penetration depth of up to 10 transport mean free paths. We develop a theoretical model which predicts the maximal-remission enh…
Optical random scattering is generally considered to be a nuisance of microscopy that limits imaging depth and spatial resolution. Wavefront shaping techniques have recently enabled optical imaging at unprecedented depth, but a remaining problem is also to attain super-resolution within complex media. To address this challenge, we introduce a new technique to focus inside of complex media by the use of a quantum reference beacon (QRB), consisting of solid-state quantum emitters with spin-dependent fluorescence. This QRB provides subwavelength guidestar feedback for wavefront shaping to achieve…
