Controlling the propagation and interaction of light in complex media has sparked major interest in the last few years. Unfortunately, spatial light modulation devices suffer from limited speed that precludes real-time applications such as imaging in live tissue. To address this critical problem we introduce a phase-control technique to characterize complex media based on the use of fast 1D spatial light modulators and a 1D-to-2D transformation performed by the same medium being analyzed. We implement the concept using a micro-electro-mechanical grating light valve (GLV) with 1088 degrees of freedom modulated at 350 KHz, enabling unprecedented high-speed wavefront measurements. We continuously measure the transmission matrix, calculate the optimal wavefront and project a focus through various dynamic scattering samples in real-time, all within 2.4 ms per cycle. These results improve prior wavefront shaping modulation speed by more than an order of magnitude and open new opportunities for optical processing using 1D-to-2D transformations.
We present an ultra-thin hybrid imaging system based on an optical multimode fiber (MMF) and an optical fiber hydrophone that combines optical resolution photoacoustic and fluorescence microscopy. To control the illumination at the distal tip of the MMF, a digital micromirror device modulates the amplitude of the optical wavefront which is coupled into the MMF. A set of pre-calibrated speckle illuminations combined with a reconstruction algorithm enables photoacoustic and fluorescence imaging of samples located at the distal tip of the fiber with optical resolution determined by the numerical aperture.Here we employ an approach that does not require focus-1 arXiv:1812.11206v1 [physics.optics]
Speckle based imaging consists of forming a superresolved reconstruction of an unknown sample from lowresolution images obtained under random inhomogeneous illuminations (speckles). In a blind context where the illuminations are unknown, we study the intrinsic capacity of speckle-based imagers to recover spatial frequencies outside the frequency support of the data, with minimal assumptions about the sample.We demonstrate that, under physically realistic conditions, the covariance of the data has a super-resolution power corresponding to the squared magnitude of the imager point spread function. This theoretical result is important for many practical imaging systems such as acoustic and electromagnetic tomographs, fluorescence and photoacoustic microscopes, or synthetic aperture radar imaging. A numerical validation is presented in the case of fluorescence microscopy.
Fluorescence Lifetime Imaging (FLIM) is an attractive microscopy method in the life sciences, yielding information on the sample otherwise unavailable through intensity-based techniques. A novel Noise-Corrected Principal Component Analysis (NC-PCA) method for time-domain FLIM data is presented here. The presence and distribution of distinct microenvironments are identified at lower photon counts than previously reported, without requiring prior knowledge of their number or of the dye's decay kinetics. A noise correction based on the Poisson statistics inherent to Time-Correlated Single Photon Counting is incorporated. The approach is validated using simulated data, and further applied to experimental FLIM data of HeLa cells stained with membrane dye di-4-ANEPPDHQ. Two distinct lipid phases were resolved in the cell membranes, and the modification of the order parameters of the plasma membrane during cholesterol depletion was also detected. Noise-corrected Principal Component Analysis of FLIM data resolves distinct microenvironments in cell membranes of live HeLa cells.
The blind structured illumination microscopy strategy proposed by Mudry et al. is fully re-founded in this paper, unveiling the central role of the sparsity of the illumination patterns in the mechanism that drives super-resolution in the method. A numerical analysis shows that the resolving power of the method can be further enhanced with optimized one-photon or two-photon speckle illuminations. A much improved numerical implementation is provided for the reconstruction problem under the image positivity constraint. This algorithm rests on a new preconditioned proximal iteration faster than existing solutions, paving the way to 3D and real-time 2D reconstruction.
We consider a fluorescence microscope in which several three-dimensional images of a sample are recorded for different speckle illuminations. We show, on synthetic data, that by summing the positive deconvolution of each speckle image, one obtains a sample reconstruction with axial and transverse resolutions that compare favorably to that of an ideal confocal microscope.
25Super-resolution fluorescence microscopy has been instrumental to progress in biology. Yet, the photo-26 induced toxicity, the loss of resolution into scattering samples or the complexity of the experimental setups 27 curtail its general use for functional cell imaging. Here, we describe a new technology for tissue imaging 28 reaching a 114nm/8Hz resolution at 30 µm depth. Random Illumination Microscopy (RIM) consists in 29 shining the sample with uncontrolled speckles and extracting a high-fidelity super-resolved image from the 30 variance of the data using a reconstruction scheme accounting for the spatial correlation of the illuminations. 31 Super-resolution unaffected by optical aberrations, undetectable phototoxicity, fast image acquisition rate and 32 ease of use, altogether, make RIM ideally suited for functional live cell imaging in situ. RIM ability to image 33 molecular and cellular processes in three dimensions and at high resolution is demonstrated in a wide range 34 of biological situations such as the motion of Myosin II minifilaments in Drosophila. 35 36 37 38
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