Abstract: A SIngle-frame LAbel-free Cell Tomography (SILACT) system and methods are provided to reconstruct 3D Refractive Index (RI) distribution of cells at over 10,000 volumes/second while resolving subcellular compartments without fluorescence labelling. The SILACT includes a high-speed interference microscope with multiplex illumination and a fast reconstruction method utilizing a pre-trained physics-incorporating Deep Neural Network (DNN). With SILACT, it is demonstrated that 3D imaging cytometry at a throughput of over 20,000 cells/second can be achieved, and transient dynamics of Red Blood Cells (RBCs) undergoing shear-induced 3D deformation inside a microfluidic channel can be observed.

Abstract: Systems and methods herein provide improved, high-throughput multiphoton imaging of thick samples with reduced emission scattering. The systems and methods use structured illumination to modify the excitation light. A reconstruction process can be applied to the resulting images to recover image information free of scattering. The disclosed systems and methods provide high throughput, high signal-to-noise ratio, and high resolution images that are depth selective.

Abstract: Since the fat content of pork is a deciding factor in grading the quality of meat, the use of a noninvasive subcutaneous probe for real-time, in situ monitoring of the fat components is of importance to vendors and other interested parties. Fortunately, in situ, in vivo monitoring of subcutaneous fat can be accomplished with spatially offset Raman spectroscopy (SORS) using a fiber-optic probe. The probe acquires Raman spectra as a function of spatial offset. These spectra are used to determine the relative composition of fat-to-skin. The Raman intensity ratio varies disproportionately depending on the fat content, with variations in slope that are correlated to the thickness of the fat layer. Ordinary least square (OLS) regression using two components indicates that depth-resolved SORS spectra reflect the relative thickness of the fat layer.

Abstract: Our high phase sensitivity wide-field phase cancellation interferometry system allows single-shot, label-free optical sensing of neural action potentials via imaging of optical path length changes. Single-shot sensing and monitoring of single neurons within a neural network should lead to a more comprehensive understanding neural network processing, which is beneficial for the advancement in the field of neuroscience as well as its biomedical applications and impact. Our system cancels the phase profile of the resting neuron from the phase profile of the spiking neuron, improving the sensitivity by two orders of magnitude. Using a detector with an extremely large well depth and an appropriately biased interferometer increases the sensitivity by another order of magnitude, yielding a measurement that is three orders of magnitude more sensitive than those possible with other microscopes.

Abstract: Since the fat content of pork is a deciding factor in grading the quality of meat, the use of a noninvasive subcutaneous probe for real-time, in situ monitoring of the fat components is of importance to vendors and other interested parties. Fortunately, in situ, in vivo monitoring of subcutaneous fat can be accomplished with spatially offset Raman spectroscopy (SORS) using a fiber-optic probe. The probe acquires Raman spectra as a function of spatial offset. These spectra are used to determine the relative composition of fat-to-skin. The Raman intensity ratio varies disproportionately depending on the fat content, with variations in slope that are correlated to the thickness of the fat layer. Ordinary least square (OLS) regression using two components indicates that depth-resolved SORS spectra reflect the relative thickness of the fat layer.

Abstract: Our high phase sensitivity wide-field phase cancellation interferometry system allows single-shot, label-free optical sensing of neural action potentials via imaging of optical path length changes. Single-shot sensing and monitoring of single neurons within a neural network should lead to a more comprehensive understanding neural network processing, which is beneficial for the advancement in the field of neuroscience as well as its biomedical applications and impact. Our system cancels the phase profile of the resting neuron from the phase profile of the spiking neuron, improving the sensitivity by two orders of magnitude. Using a detector with an extremely large well depth and an appropriately biased interferometer increases the sensitivity by another order of magnitude, yielding a measurement that is three orders of magnitude more sensitive than those possible with other microscopes.

Abstract: Noninvasive glucose monitoring has been a long-standing need in diabetes management. Among many approaches to meeting this need, Raman spectroscopy has attracted attention due to its molecular specificity. Previous Raman-based glucose sensing can predict blood glucose concentration based on a statistical correlation between the reference glucose concentration and unspecified spectral features. However, the lack of glucose Raman peaks and non-prospective prediction have led to questions about the effectiveness of in vivo Raman spectroscopy for transcutaneous glucose sensing. Here, we disclose technology for directly observing distinct glucose Raman spectra from skin. The Raman signal intensities were proportional to the reference glucose concentrations in three live swine glucose clamping experiments. Tracking the spectral intensity based on the linearity enables prospective prediction with high accuracy in within-subject and inter-subject models.

Abstract: Systems and methods herein provide improved, high-throughput multiphoton imaging of thick samples with reduced emission scattering. The systems and methods use structured illumination to modify the excitation light. A reconstruction process can be applied to the resulting images to recover image information free of scattering. The disclosed systems and methods provide high throughput, high signal-to-noise ratio, and high resolution images that are depth selective.

Abstract: Refractive index of biological specimens is a source of intrinsic contrast that can be explored without any concerns of photobleaching or harmful effects caused by extra contrast agents. This feature also contains rich information that can be related to the metabolism of cells at the cellular and subcellular levels. The present invention relates to systems and methods that can provide, without any moving parts, the 3-D refractive index map of continuously flowing biological samples in a micro-fluidic channel, for example.

Abstract: In the systems and methods of the present invention a multifocal multiphoton imaging system has a signal to noise ratio (SNR) that is reduced by over an order of magnitude at imaging depth equal to twice the mean free path scattering length of the specimen. An MMM system based on an area detector such as a multianode photomultiplier tube (MAPMT) that is optimized for high-speed tissue imaging. The specimen is raster-scanned with an array of excitation light beams. The emission photons from the array of excitation foci are collected simultaneously by a MAPMT and the signals from each anode are detected using high sensitivity, low noise single photon counting circuits. An image is formed by the temporal encoding of the integrated signal with a raster scanning pattern. A deconvolution procedure taking account of the spatial distribution and the raster temporal encoding of collected photons can be used to improve decay coefficient.

Abstract: In the systems and methods of the present invention a multifocal multiphoton imaging system has a signal to noise ratio (SNR) that is reduced by over an order of magnitude at imaging depth equal to twice the mean free path scattering length of the specimen. An MMM system based on an area detector such as a multianode photomultiplier tube (MAPMT) that is optimized for high-speed tissue imaging. The specimen is raster-scanned with an array of excitation light beams. The emission photons from the array of excitation foci are collected simultaneously by a MAPMT and the signals from each anode are detected using high sensitivity, low noise single photon counting circuits. An image is formed by the temporal encoding of the integrated signal with a raster scanning pattern. A deconvolution procedure taking account of the spatial distribution and the raster temporal encoding of collected photons can be used to improve decay coefficient.

Abstract: An imaging system is provided that includes a pulsed light source providing pulsed light and is applicable to both microscopes and endoscopes. One or more optical elements with certain dispersive properties are positioned to receive the pulsed light and apply selective dispersive properties to shift the focal plane according to the user and to produce two photon (2p) wide field uniform illumination and 2p wide field structured illumination for the purpose of improving the optical axial resolution and rejection of background signal. An imaging element receives the signal arising from the 2p wide field uniform illumination and 2p wide field structured illumination and produces a respective 3D resolved image of a sample.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Abstract: Refractive index of biological specimens is a source of intrinsic contrast that can be explored without any concerns of photobleaching or harmful effects caused by extra contrast agents. This feature also contains rich information that can be related to the metabolism of cells at the cellular and subcellular levels. The present invention relates to systems and methods that can provide, without any moving parts, the 3-D refractive index map of continuously flowing biological samples in a micro-fluidic channel, for example.

Abstract: An imaging system is provided that includes a pulsed light source providing pulsed light and is applicable to both microscopes and endoscopes. One or more optical elements with certain dispersive properties are positioned to receive the pulsed light and apply selective dispersive properties to shift the focal plane according to the user and to produce two photon (2p) wide field uniform illumination and 2p wide field structured illumination for the purpose of improving the optical axial resolution and rejection of background signal. An imaging element receives the signal arising from the 2p wide field uniform illumination and 2p wide field structured illumination and produces a respective 3D resolved image of a sample.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Abstract: An imaging system is provided that includes a optical pulse generator for providing an optical pulse having a spectral bandwidth and includes monochromatic waves having different wavelengths. A dispersive element receives a second optical pulse associated with the optical pulse and disperses the second optical pulse at different angles on the surface of the dispersive element depending on wavelength. One or more focal elements receives the dispersed second optical pulse produced on the dispersive element. The one or more focal element recombine the dispersed second optical pulse at a focal plane on a specimen where the width of the optical pulse is restored at the focal plane.

Abstract: An imaging system is provided that includes a optical pulse generator for providing an optical pulse having a spectral bandwidth and includes monochiromatic waves having different wavelengths. A dispersive element receives a second optical pulse associated with the optical pulse and disperses the second optical pulse at different angles on the surface of the dispersive element depending on wavelength. One or more focal elements receives the dispersed second optical pulse produced on the dispersive element. The one or more focal element recombine the dispersed second optical pulse at a focal plane on a specimen where the width of the optical pulse is restored at the focal plane.