A microscopy apparatus comprises a microscope comprising a stage configured to hold a tissue sample, a UV laser assembly configured to emit a UV laser beam to a viewing area of the tissue sample, and an IR laser assembly configured to emit an IR laser beam to the viewing area of the tissue sample. The UV and IR laser assemblies are oriented so as to emit the respective UV and IR laser beams in a same direction.

Systems for operating a microfluidic device are described. The systems comprise a first surface configured to interface and operatively couple with a microfluidic device and a lid configured to retain the microfluidic device on the first surface. The lid comprises a first portion having a first fluid port configured to operatively couple with and flow fluidic medium into and/or out of a first fluid inlet/outlet of the microfluidic device and a second portion having a second fluid port configured to operatively couple with and flow fluidic medium into and/or out of a second fluid inlet/outlet of the microfluidic device. The second portion of the lid is separable from the first portion and movable between a closed position in which the second fluid port of the second portion of the cover is operatively coupled with the second fluid inlet/outlet of the microfluidic device and an open position in which a portion of the microfluidic device that contains the second fluid inlet/outlet is exposed. Other embodiments are described.

Deterioration of optical characteristics is suppressed. An optical measurement device according to an embodiment includes: an excitation light source (101 to 103) that emits excitation light having a wavelength of at least 450 nanometers or less; a lens structure (116) that condenses the excitation light at a predetermined position; a fluorescence detection system (140) that detects fluorescence emitted from a particle by excitation of the particle present at the predetermined position by the excitation light; and a scattered light detection system (130) that detects scattered light generated by the excitation light being scattered by the particle present at the predetermined position, and the lens structure includes a plurality of lenses (21, 22, 23, 25, 26, 28) arranged along an optical axis of the excitation light and a lens frame (10) that holds the plurality of lenses, and a position of at least one of the plurality of lenses in the lens frame is determined by abutting on a lens adjacent to the lens.

A microscopy imaging system and methods for improving an assaying of a sample is disclosed.

This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

A photon peak event detection system accepts an analog output from a photon sensor, directly digitizes the analogy output and includes a graphics processing unit (GPU) programmed to conduct a photon peak event detection in real-time via a photon count program that analyzes the digitized photon sensor output in sampling periods each having at least three consecutive data points to determine a local maximum among the consecutive data points and compare the local maximum to one or more predetermined thresholds to determine whether or not a photon was received in each sampling period, the algorithm providing photon counts to a phasor analysis program in the GPU. The phasor analysis program calculates pixelwise fluorescence lifetime phasor data in real-time and sends the data to a central processing unit.

A photon peak event detection system accepts an analog output from a photon sensor, directly digitizes the analogy output and includes a graphics processing unit (GPU) programmed to conduct a photon peak event detection in real-time via a photon count program that analyzes the digitized photon sensor output in sampling periods each having at least three consecutive data points to determine a local maximum among the consecutive data points and compare the local maximum to one or more predetermined thresholds to determine whether or not a photon was received in each sampling period, the algorithm providing photon counts to a phasor analysis program in the GPU. The phasor analysis program calculates pixelwise fluorescence lifetime phasor data in real-time and sends the data to a central processing unit.

Implementations of the disclosure are directed to predicting structured illumination parameters for a particular point in time, space, and/or temperature using estimates of structured illumination parameters obtained from structured illumination images captured by a structured illumination system. Particular implementations are directed to predicting structured illumination frequency, phase, orientation, and/or modulation order parameters.

An optical system is presented for optically imaging a sample including a nanoscale object. The optical system includes an imaging lens, an illumination source configured to provide an excitation light, a detector and a substrate for supporting the sample. A sample interface, arranged to reflect the excitation light, is formed between the sample and a first side of the substrate facing the sample when the sample is applied on the substrate. The optical imaging system is arranged such that the excitation light is sent into the substrate via the imaging lens and such that the detector receives a reference light and a scattered light. The reference light comprises a part of the excitation light reflected at the sample interface and collected by the imaging lens and the scattered light comprises a part of the excitation light scattered by the nanoscale object and collected by the imaging lens. The optical system is configured such that the nanoscale object is imaged at the detector, in response to the excitation light, by an optical contrast of an interference pattern between the reference light and the scattered light. The substrate comprises an optical coating disposed on the first side of the substrate such that the sample is in contact with the optical coating when the sample is applied on the substrate. A degree of reflection of the excitation light at the sample interface is such that the optical contrast is larger compared to the optical contrast obtained with the sample interface formed without the optical coating.