An in-line holographic microscope can be used to analyze on a frame-by-frame basis a video stream to track individual colloidal particles' three-dimensional motions. The system and method can provide real time nanometer resolution, and simultaneously measure particle sizes and refractive indexes. Through a combination of applying a combination of Lorenz-Mie analysis with selected hardware and software methods, this analysis can be carried out in near real time. An efficient particle identification methodology automates initial position estimation with sufficient accuracy to enable unattended holographic tracking and characterization.

A method of analyzing a sample receiving a particle of interest, including: defining a reference point located on a first interface of the sample, or at a known distance from the sample, along the optical axis of the optical system; acquiring a reference image transmission of the sample, the object plane of the optical system being located at a known distance from the reference point along an axis parallel to the optical axis of the optical system, and the particle of interest being located outside of the object plane; using the reference image, digitally constructing a series of reconstructed images, each associated with a predetermined offset of the object plane along the optical axis of the optical system; and using the series of reconstructed images, determining the distance along an axis parallel to the optical axis of the optical system, between the particle of interest and the reference point.

A method for 3D imaging of a sample, in a vessel having a longitudinal axis orthogonal to a horizontal plane, includes capturing, by at least three cameras located at different positions around the vessel, respective 2D images of the sample. Each image comprises pixels having associated pixel values. The optical axis of a first camera is inclined or declined at a first angle relative to the horizontal plane, with the first angle being greater than or equal to zero degrees. The optical axis of a second camera is inclined or declined at a second, larger angle relative to the horizontal plane. The method also includes generating a 3D image of the sample based on the pixel values associated with the 2D image pixels, and one or more look-up tables that collectively indicate, for pixels in each image, expected paths for light traversing the vessel and the sample.

The present disclosure provides systems and methods for sorting a cell. The system may comprise a flow channel configured to transport a cell through the channel. The system may comprise an imaging device configured to capture an image of the cell from a plurality of different angles as the cell is transported through the flow channel. The system may comprise a processor configured to analyze the image using a deep learning algorithm to enable sorting of the cell.

The present invention discloses a three-dimensional diffraction tomography microscopy imaging method based on LED array coded illumination. Firstly, acquiring the raw intensity images, three sets of intensity image stacks are acquired at different out-of-focus positions by moving the stage or using electrically tunable lens. And then, after acquiring the intensity image stacks of the object to be measured at different out-of-focus positions, the three-dimensional phase transfer function of the microscopy imaging system with arbitrary shape illumination is derived. Further, the three-dimensional phase transfer function of the microscopic system under circular and annular illumination with different coherence coefficients is obtained as well, and the three-dimensional quantitative refractive index is reconstructed by inverse Fourier transform of the three-dimensional scattering potential function. The scattering potential function is converted into the refractive index distribution. Thus, the quantitative three-dimensional refractive index distribution of the test object is obtained. The invention realizes high-resolution and high signal-to-noise ratio 3D diffraction tomography microscopic imaging of cells, tiny biological tissues and other samples.

Disclosed are methods, devices, systems and applications for camera-less, high-throughput three-dimensional imaging of particles in motion. In some aspects, a system includes a particle motion device to allow particles to move along a travel path; an optical illumination system to produce an asymmetric illumination area of light in a region of the travel path of a particle that scans over a plurality of sections of the particle at multiple time points while the particle is moving; an optical detection system optically interfaced with the particle motion device to obtain optical signal data associated with different parts of the particle corresponding to the particle's volume during motion in the travel path; and a data processing unit to process the optical signal data obtained by the optical detection system and produce data including information indicative of 3D features of the particle.

An image sensor and well structure associated with and extending away from the surface of the image sensor are provided in various apparatuses, methods, and systems for determining the position of a light emitter located in object space. An exemplary method includes (i) providing the image sensor and structure associated therewith, the structure defining a field of view for each pixel within the array of pixels; (ii) determining a light intensity value for photoactivated pixels receiving light from the light emitter; (iii) identifying a first photoactivated pixel having a local maximum of light intensity; (iv) calculating a perpendicular distance between the first photoactivated pixel and the light emitter; and (v) constructing the position of the light emitter based on a position of the first photoactivated pixel in the array of pixels and the perpendicular distance between the first photoactivated pixel and the light emitter.

A device includes a filter that enhances a beam profile of a received pulsed laser; a first optical element to direct the pulsed laser as a reference wave towards an optical sensor; an open cavity positioned between the first optical element and the optical sensor. The open cavity receives an aerosol particle, which enters the open cavity from any direction. The reference wave illuminates the aerosol particle. An illuminated particle generates and directs an object wave towards the optical sensor. A pixel array is connected to the optical sensor. The pixel array receives the reference wave and the object wave. The optical sensor creates a contrast hologram comprising an interference pattern of the illuminated particle. A processor creates an image of the illuminated particle based on the contrast hologram.

The present invention provides various methods for easily assessing cell quality of a cell production process, suitably using non-invasive visual methods and neural networks for generating predictive fluorescence images of cells to assess quality attributes. Also provided are systems and methods for carrying out such processes.

An observation container (10) includes: a bottom portion that includes a bottom wall (12A) (first plate part) and a bottom wall (12B) (second plate part) which intersect each other and that is configured to accommodate a liquid sample O as a sample containing microparticles to be imaged by imaging units (30A) and (30B) serving as an imaging device; and a region that has transparency with respect to a wavelength of light used for observation of the microparticles.