The present disclosure relates to spatially modulating the light source used in microscopy. In some cases, a light source projects a sequence of two-dimensional spatial patterns onto a sample using a spatial light modulator. In some cases, the spatial patterns are based on Hadamard matrices. In some cases, an imaging device captures frames of image data in response to light emitted by the sample and orthogonal components of the image data are analyzed by cross-correlating the image data with the spatial pattern associated with each frame. A microscope may be calibrated by illuminating a sample with the sequence of spatial patterns, capturing image data, and storing calibration that maps each pixel of the spatial light modulator to at least one pixel of the imaging device.

An assembly for measurements of one or more optical parameters of a medium is disclosed. The assembly comprises a light sheet generator, a light intensity modulator, a holder for a sample, and an optical sensor.

The light sheet generator is configured to provide a polychromatic light sheet comprising a light spectrum extending in a first spatial dimension, wherein the polychromatic light sheet has a propagation path in a second spatial dimension.

The light sheet generator comprises a light source configured to provide white light, a dispersive element configured to spread the white light in the first spatial dimension to provide the light spectrum, and an optical slit extending in the first spatial dimension configured to provide the polychromatic light sheet by limitation of the spread white light.

The light intensity modulator is configured to provide an intensity modulated polychromatic light sheet by applying—to the polychromatic light sheet—an intensity modulation having a periodical (or substantially periodical) pattern in the first spatial dimension.

The holder for a sample of the medium is configured to enable the intensity modulated polychromatic light sheet to illuminate the sample.

The optical sensor is configured to record intensity of light exiting the sample over the light spectrum for provision of the one or more optical parameters.

A method of using the assembly for measuring one or more optical parameters of a medium is also disclosed.

An assembly for measurements of one or more optical parameters of a medium is disclosed. The assembly comprises a light sheet generator, a light intensity modulator, a holder for a sample, and an optical sensor. A method of using the assembly for measuring one or more optical parameters of a medium is also disclosed.

Certain disclosed embodiments concern an integrated imaging system that combined light-sheet microscopy, which enables considerable speed and phototoxicity gains, with quantitative-phase imaging. A method for using such imaging systems also is disclosed. In an exemplary embodiment, an integrated imaging system was used for multivariate investigation of live-cells in microfluidics.

A system for processing Raman scattering light from a sample is provided. The system includes a source, a digital mirror device (DMD), a detector, and an analyzer. The DMD is configured to reflect Raman scattering light and includes micromirrors selectively controllable between ON and OFF states. The detector is configured to detect Raman scattering light and to produce signals representative of the Raman scattering light. The analyzer is in communication with the light source, the DMD, the detector, and a memory storing instructions, which instructions when executed cause the processor to: a) control the light source to produce a beam of light for interrogating the sample; b) control the DMD to place in an ON or OFF state based on one or more known spectral shapes stored in the memory; and c) process the Raman scattering light reflected by the micromirrors in the ON state.

A time-resolved hyper-spectral imaging system for imaging a sample, includes a radiation source suitable for illuminating the sample repeatably, a first optical system configured to form an image I of the sample on a spatial light modulator forming a transmission or reflection mask P, a processor connected to the spatial light modulator and configured to make the transmission or reflection mask P vary for each repetition of the illumination, a second optical system suitable for focusing the radiation transmitted or reflected by the spatial light modulator so as to form, in its image focal plane, a partial image S=P.Math.I; the imaging system being wherein it comprises: a dispersive device comprising a slit placed in the image focal plane of the second optical system, the dispersive device being suitable for spatially splitting the various wavelengths of the radiation transmitted or reflected by the spatial light modulator; a streak camera arranged so as to be illuminated by the radiation issuing from the dispersive device and configured to acquire a plurality of time-resolved partial images of the sample, the images being associated with respective and different transmission or reflection masks P, the streak camera being connected to the processor and the processor also being configured to combine the partial images of the sample so as to construct a 4D image cube I.sub.tot forming an image resolved in time and in wavelength of the sample; and corresponding time-resolved hyper-spectral imaging method for imaging a sample.

An optical detection system includes a sample portion accommodating a sample, a wave source emitting waves to the sample portion, an optical portion provided on a path of an output wave output from the sample portion, and comprising a first spatial light modulator that modulates part of the output wave to a first wave and a second spatial light modulator that modulates part of the output wave to a second wave, a lens portion focusing the first wave and the second wave output from the optical portion, and a detection portion detecting a focused wave that is focused by the lens portion, in which the first spatial light modulator and the second spatial light modulator modulate the output wave such that the first wave and the second wave have destructive interference with respect to the sample under an already known condition.

A system, including an optical imaging assembly configured to image a sample at an object plane to an image plane; an image sensor arranged at the image plane and configured to capture images of the sample for a field of view of the system; a light source configured to emit light having a wavelength, λ; a spatial light modulator (SLM) arranged to receive the light emitted from the light source and to provide a spatially modulated light pattern; one or more optical elements arranged to receive the spatially modulated light pattern from the SLM and to direct the spatially modulated light pattern to the image plane; and an electronic controller in communication with the image sensor and the spatial light modulator, the electronic controller being programmed to identify one or more targets in the field of view of the optical imaging assembly and to control the spatial light modulator to selectively direct light from the light source to the one or more targets identified by the electronic controller.

Methods, systems and kits are described herein for detecting the results of an assay. In particular, the methods, systems and devices of the present disclosure rely on a difference between the diffusion rates of a reporter molecule and an analyte of interest in order to quantify an amount of analyte in a microfluidic device. The analyte may be a secreted product of a biological micro-object.

Lightening method of at least one biological sample (S), in which said at least one biological sample includes at least one or more fluorophores, at the focal point (F) of at least one objective lens (L) having a main optical axis (z-z), the method comprising the following operational steps:—lightening (step 10) said at least one biological sample (S) with at least one excitation beam (EB), which propagates between said at least one objective lens (L) and said at least one biological sample (S) along at least one first propagation axis (a-a);—lightening (step 20) said at least one biological sample (S) with at least two depletion beams (DB, DB′), which propagate between said at least one objective lens (L) and said at least one biological sample (S) along the respective second propagation axes (b-b, said depletion beams being donut-shaped, each one in a plane orthogonal to the respective second propagation axis (b-b, b′-b′); whereby said at least one first propagation axis (a-a) and said at least second propagation axes (b-b, b′-b′) are angularly inclined with each other, and said at least one first propagation axis (a-a) and said second propagation axes (b-b, b′-b′) intersect on said at least one biological sample (s) only at the focal point (F) of said at least one objective lens (L), so that an effective fluorescence volume (FV) is generated in said at least one biological sample (S) which is limited both orthogonally and axially with respect to said main optical axis (z-z).