A method for generating a three-dimensional luminescence image includes setting a focal interval between two-dimensional images in accordance with localization of luminescence in a three-dimensional sample. The three-dimensional sample contains a plurality of cells prepared to be luminescent and has a three-dimensional shape. The two-dimensional images have mutually different focal planes and are acquired at the focal interval. The method further includes acquiring a two-dimensional image set including two-dimensional images at the focal interval that is set by imaging the three-dimensional sample under an unirradiated condition; and generating a three-dimensional luminescence image by combining the two-dimensional images included in the two-dimensional image set together.

Systems and methods in accordance with various embodiments of the invention are capable of rapid analysis and classification of cellular samples based on cytomorphological properties. In several embodiments, cells suspended in a fluid medium are passed through a microfluidic channel, where they are focused to a single stream line and imaged continuously. In a number of embodiments, the microfluidic channel establishes flow that enables individual cells to each be imaged at multiple angles in a short amount of time. A pattern recognition system can analyze the data captured from high-speed images of cells flowing through this system and classify target cells. In this way, the automated platform creates new possibilities for a wide range of research and clinical applications such as (but not limited to) point of care services.

In order to simply and accurately acquire a three-dimensional image of a sample to observe the sample, a light-sheet microscope according to the present invention includes: a duct in which the sample can flow; a syringe pump for moving the sample in the duct; a cylindrical lens for causing planar illumination light along a plane intersecting the direction in which the sample is moved by the syringe pump to enter the duct; an objective lens that is disposed so as to face the radiation plane of the illumination light caused by the cylindrical lens to enter the duct and that collects fluorescence emitted from the sample moved by the syringe pump so as to pass through the radiation plane of the illumination light; and a camera for acquiring an image of the fluorescence from the sample collected by the objective lens.

The invention concerns a method for identifying biological particles from a stack of holographic images obtained by means of an optical system. A stack of image blocks centred on the biological particle to be analysed is extracted from the stack of images and a reference block corresponding to the focus plan is determined. A characteristic magnitude is calculated for each block of the stack and the profile of this characteristic magnitude along the optical axis of the system is compared with a plurality of standard profiles relative to known types of particle. Alternatively, blocks of the stack are extracted from the stack of blocks for predetermined defocusing deviations and the extracted blocks are compared with standard blocks relative to known types of particle.

Holograms of colloidal dispersions encode comprehensive information about individual particles' three-dimensional positions, sizes and optical properties. Extracting that information typically is computation-ally intensive, and thus slow. Machine-learning techniques based on support vector machines (SVMs) can analyze holographic video microscopy data in real time on low-power computers. The resulting stream of precise particle-resolved tracking and characterization data provides unparalleled insights into the composition and dynamics of colloidal dispersions and enables applications ranging from basic research to process control and quality assurance.

Methods, devices, and systems for performing digital assays are provided. In certain aspects, the methods, devices, and systems can be used for the amplification and detection of nucleic acids. In certain aspects, the methods, devices, and systems can be used for the recognition, detection, and sizing of droplets in a volume. Also provided are compositions and kits suitable for use with the methods and devices of the present disclosure.

A method for enhancing gating performance of a cell sorter to prepare an enriched specimen for optical tomography cell analysis includes introducing a specimen into a FACS to generate 2D event data; generating a first scatterplot of the 2D data; identifying target objects; constructing a boundary within the first scatterplot to produce a first gate; counting target objects within the first gate; comparing the number of target objects within the first gate to a first predetermined value and adjusting the first gate as necessary. A boundary around a set of target objects is constructed in a second scatterplot to produce a subset second gate and target objects within the second gate are counted and the count compared to a second predetermined value. When a boundary around target objects meets specifications the first and second gates are stored in memory and used to enrich patient specimens.

The present disclosure relates to systems and methods for cellular imaging and identification through the use of a light sheet flow cytometer. In one implementation, a light sheet flow cytometer may include a light source configured to emit light having one or more wavelengths, at least one optical element configured to form a light sheet from the emitted light, a microfluidic channel configured to hold a sample, and an imaging device. The imaging device may be adapted to forming 3-D images of the sample such that identification tags attached to the sample are visible.

With a water and debris mixture, a method of quantifying debris content may include obtaining at least one image of a sample of the water and debris mixture. The image may be analyzed to quantify the debris content.

A method for lens-free imaging of a sample or objects within the sample uses multi-height iterative phase retrieval and rotational field transformations to perform wide FOV imaging of pathology samples with clinically comparable image quality to a benchtop lens-based microscope. The solution of the transport-of-intensity (TIE) equation is used as an initial guess in the phase recovery process to speed the image recovery process. The holographically reconstructed image can be digitally focused at any depth within the object FOV (after image capture) without the need for any focus adjustment, and is also digitally corrected for artifacts arising from uncontrolled tilting and height variations between the sample and sensor planes. In an alternative embodiment, a synthetic aperture approach is used with multi-angle iterative phase retrieval to perform wide FOV imaging of pathology samples and increase the effective numerical aperture of the image.