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.

A method for measuring a position of a fluorophore includes configuring a set of compact light distributions, the set having at least one member, each light distribution characterized by a center, so that there is substantially zero intensity at the center of the set of compact light distributions. The method additionally includes moving the set of compact light distributions in relation to a set of hypothesized positions of the fluorophore, detecting, in a plurality of locations corresponding to the hypothesized set of positions, a set of images; and estimating the position of the fluorophore, by determining from the set of images a set of parameters describing the position of the fluorophore using an inverse problem method.

Among other things, an imaging device has a photosensitive array of pixels, and a surface associated with the array is configured to receive a specimen with at least a part of the specimen at a distance from the surface equivalent to less than about half of an average width of the pixels.

The disclosure provides a method of generating an artificial fluorescent image of cells is provided. The method includes receiving a brightfield image generated by a brightfield microscopy imaging modality of at least a portion of cells included in a specimen, applying, to the brightfield image, at least one trained model, the trained model being trained to generate the artificial fluorescent image based on the brightfield image, receiving the artificial fluorescent image from the trained model

A MEM-based device and method of fabrication, the device comprising a biochip substrate comprising one or more compliant materials, a plurality of mechanical and electrical micro-sensors configured in an array to simultaneously measure electrical and mechanical properties of a sample, wherein a first mechanical micro-sensor is formed as a patterned layer of at least one of the compliant materials, wherein the patterned layer is coupled to a first pillar comprising a dielectric material formed onto the compliant materials, the first pillar being coated with a metal film at a contact surface with the sample and along a side of the first pillar to act as a conductive probe for the first electrical micro-sensor, and wherein the first pillar is formed on the first mechanical micro-sensor to transfer a force to the first mechanical micro-sensor.

Multiplexed fluorescent imaging which is essential for finding out how various biomolecules are spatially distributed in cells or tissues is disclosed. The present disclosure may obtain 10 or more different biomolecule images with one labeling and imaging by newly designing selection of fluorophores, detection spectral ranges, and signal unmixing algorithm. The present disclosure is a blind unmixing technology for unmixing an image without an emission spectrum of fluorophore, and in this technology, 4 pairs of fluorophores are used, and each pair consists of two fluorophores in which emission spectra are overlapped. Each pair of fluorophores is strongly excited by only one excitation laser. Two images with different detection spectral ranges are obtained for each pair, and two images are unmixed via mutual information minimization without fluorophore emission spectrum information. Two images also may be unmixed via Gram-Schmidt orthogonalization and fluorescence measurement based unmixing. This signal unmixing is repeated for each pair of fluorophores. Furthermore, a total of 10 or more fluorophores may be simultaneously used by adding two large stoke's shift fluorophores emitting light in wavelength ranges that does not overlap with the emission spectra of the above 8 fluorophores.

A light source unit includes: a housing; a semiconductor laser that is disposed in the housing and that radiates excitation light; a first condenser optical system that condenses the excitation light; a dichroic mirror that selectively reflects the excitation light; a second condenser optical system that condenses the excitation light; a wavelength conversion member that performs wavelength conversion of the excitation light and emits wavelength-converted light; an emission section that outputs the wavelength-converted light transmitted through the second condenser optical system and the dichroic mirror; and a light blocking section that is disposed between an inner surface of the housing, the inner surface being in a traveling direction of the excitation light toward a reflection surface of the dichroic mirror, and a back surface, the back surface being an opposite side of the reflection surface, or is disposed on the inner surface of the housing.

The present invention is to provide methods and devices that monitoring health and diagnosing a disease by directly measuring the biomarkers inside a cell (intra-cellular detection) rapidly and easily.

Methods and systems are provided for improving resolution, acquisition speed, and/or illumination dose for microscopy systems. In some embodiments, a microscopy system having multiple objective setups may include illumination generators to provide selectively-blanked illumination line scans, objective lenses to introduce the selectively-blanked illumination line scans to a sample and to collect fluorescence emissions from the sample, and detectors to receive the fluorescence emissions from the objective lenses. The microscopy system may also include one or more processors in operative communication with the detectors, which may combine the fluorescence emissions to generate a composite image.

In some aspects, the present disclosure relates to methods for reducing the crowding of signals, for example optical crowding, that can occur when nucleic acids are detected in a sample in multiplex, which can make it difficult to resolve individual signals and can lead to a reduced dynamic range. In some aspects, the present disclosure relates to methods for reducing signal crowding in the detection of multiple target nucleic acid sequences in a sample, e.g., using hybridization probes, wherein signal crowding from said hybridization probes is reduced. The methods herein have particular applicability in the detection of barcode sequences by sequencing-by-hybridization (SBH) methods, including those relying on combinatorial labelling schemes and decoding of the barcodes by sequential cycles of decoding using hybridization probes. Also provided are kits comprising probes for use in such methods.