Methods for producing a high resolution image of a sample are provided. In some embodiments, the method includes: detecting first and second sets of spatially-dependent emission signals from a sample labeled with a fluorescent polymeric dye; and producing a high resolution fluorescence image of the sample from the detected first and second sets of spatially-dependent emission signals. In some embodiments, the sample is a cell. Also provided are systems for imaging a sample that include a high resolution light microscope including a light source configured to irradiate a field of view with an excitation light; a photodetector configured to detect an emission signal: and a polarization modulator disposed in the light pathway between the light source and the photodetector; and a sample labelled with a polymeric dye and disposed in the field of view.

An imaging system for imaging biological samples includes at least one main channel including at least one imaging space, the at least one main channel configured to transport the samples in a fluid, at least one reorientation unit configured to manipulate an orientation of the samples in the fluid, and at least one imaging unit configured to receive detection light emitted by the samples in the at least one imaging space.

An analyzer of a component in a sample fluid includes an optical source and an optical detector defining a beam path of a beam, wherein the optical source emits the beam and the optical detector measures the beam after partial absorption by the sample fluid, a fluid flow cell disposed on the beam path defining an interrogation region in the a fluid flow cell in which the optical beam interacts with the sample fluid and a reference fluid; and wherein the sample fluid and the reference fluid are in laminar flow, and a scanning system that scans the beam relative to the laminar flow within the fluid flow cell, wherein the scanning system scans the beam relative to both the sample fluid and the reference fluid.

Microfluidic devices, systems and techniques in connection with particle sorting in liquid, including cytometry devices and techniques and applications in chemical or biological testing and diagnostic measurements.

A mask structure optimization device includes a classification target image size acquisition unit that is configured to acquire a size of a classification target image which is an image including a classification target, a mask size setting unit that is configured to set a size of a mask applied to the classification target image, a brightness detection unit that is configured to detect a brightness of each pixel within the classification target image at a position on an opposite side of the mask from the classification target image, a sum total brightness calculation unit that is configured to calculate the sum total brightness of the each pixel within the classification target image detected by the brightness detection unit, an initial value setting unit that is configured to set an initial value for a mask pattern of the mask, and a movement unit that is configured to relatively move the mask with respect to the classification target image. The sum total brightness calculation unit is configured to calculate the sum total brightness of the each pixel within the classification target image every time the movement unit relatively moves the mask by a predetermined movement amount. The mask structure optimization device further includes a mask pattern optimization unit that is configured to optimize the mask pattern of the mask on the basis of the sum total brightness.

Microfluidic devices, systems and techniques in connection with particle sorting in liquid, including cytometry devices and techniques and applications in chemical or biological testing and diagnostic measurements.

Disclosed is a method for detecting nano-particles, comprising the steps of (1) compressing a sample liquid to be tested into a sample liquid flow by hydrodynamic focusing using a sheath fluid; (2) irradiating a measuring light to the sample liquid flow, wherein a single nano-particle in the sample liquid flow is irradiated by the measuring light for a duration of 0.1-10 milliseconds; (3) defining the area in which the measuring light irradiates the sample liquid flow as a detecting area, and collecting light signals emitted from the area irradiated by the measuring light by a lens imaging system, and allowing the light signals collected by the lens imaging system to pass a field stop, so as to filter out the light signals outside the detecting area and enrich the light signals from the detecting area; and (4) subjecting the light signals enriched by the field stop to optoelectronic signal conversion. The method can achieve detection for nano-particles with a low refractive index and a particle size of 24-1000 nm as well as nano-scale gold particles with a particle size of 6.7-150 nm.

Methods, systems, and devices are disclosed for imaging particles and/or cells using flow cytometry. In one aspect, a method includes transmitting a light beam at a fluidic channel carrying a fluid sample containing particles; optically encoding scattered or fluorescently-emitted light at a spatial optical filter, the spatial optical filter including a surface having a plurality of apertures arranged in a pattern along a transverse direction opposite to particle flow and a longitudinal direction parallel to particle flow, such that different portions of a particle flowing over the pattern of the apertures pass different apertures at different times and scatter the light beam or emit fluorescent light at locations associated with the apertures; and producing image data associated with the particle flowing through the fluidic channel based on the encoded optical signal, in which the produced image data includes information of a physical characteristic of the particle.

In one aspect, the present teachings provide a system for performing cytometry that can be operated in three operational modes. In one operational mode, a fluorescence image of a sample is obtained by exciting one or more fluorophore(s) present in the sample by an excitation beam formed as a superposition of a top-hat-shaped beam with a plurality of beams that are radiofrequency shifted relative to one another. In another operational mode, a sample can be illuminated successively over a time interval by a laser beam at a plurality of excitation frequencies in a scanning fashion. The fluorescence emission from the sample can be detected and analyzed, e.g., to generate a fluorescence image of the sample. In yet another operational mode, the system can be operated to illuminate a plurality of locations of a sample concurrently by a single excitation frequency, which can be generated, e.g., by shifting the central frequency of a laser beam by a radiofrequency. For example, a horizontal extent of the sample can be illuminated by a laser beam at a single excitation frequency. The detected fluorescence radiation can be used to analyze the fluorescence content of the sample, e.g., a cell/particle.

In one aspect, a method of sorting cells in a flow cytometry system is disclosed, which includes illuminating a cell with radiation having at least two optical frequencies shifted from one another by a radiofrequency to elicit fluorescent radiation from the cell, detecting the fluorescent radiation to generate temporal fluorescence data, and processing the temporal fluorescence data to arrive at a sorting decision regarding the cell without generating an image (i.e., a pixel-by-pixel image) of the cell based on the fluorescence data. In other words, while the fluorescence data can contain image data that would allow generating a pixel-by-pixel fluorescence intensity map, the method arrives at the sorting decision without generating such a map. In some cases, the sorting decision can be made with a latency less than about 100 microseconds. In some embodiments, the above method of sorting cells can have a sub-cellular resolution, e.g., the sorting decision can be based on characteristics of a component of the cell. In some embodiments in which more than two frequency-shifted optical frequencies are employed, a single radiofrequency shift is employed to separate the optical frequencies while in other such embodiments a plurality of different radiofrequency shifts are employed.