Abstract: Provided is a method of quantifying a target substance present in a solvent comprises acquiring a first fluorescence decay curve and a second fluorescence decay curve to acquire some factors from the curves.

Abstract: Systems and methods to identify and/or reduce or eliminate sample motion artifacts are disclosed. Sample motion artifacts may be reduced or eliminated using scan patterns where an acquisition time difference between when perimeter pixels in adjacent tiles are acquired is reduced, as compared to a conventional raster scan to reduce or eliminate discontinuities that would otherwise appear at tile boundaries in an image. In some embodiments, test images acquired using relatively small test scan patterns or intensities of test points acquired at different times may be compared to determine whether sample motion has occurred. In some embodiments, intensity of adjacent pixels at a tile boundary are compared. In some embodiments, intensity of one or more single pixels is monitored over time to determine whether sample motion has occurred over a period of time. In some embodiments, a flattening or reshaping tool may be used to suppress sample motion during imaging.

Abstract: The present disclosure provides a measurement system and a measurement method related to the field of measurement techniques, the measurement system comprising: a light source configured to produce an original beam, wherein a return beam is formed by the original beam returning from a measured area of a measured object; an optical assembly configured to obtain a pending beam based on the return beam, wherein at least part of the pending beam acts as a first beam; a first detection device configured to obtain first detection information based on the first beam; a mobile device configured to move the optical assembly and the measured object with respect to each other in a direction of an optical axis of the optical assembly; and a processing system configured to determine an actual distance between the optical assembly and a fixed plane of the measured object at each first moment according to the first detection information at each first moment of a plurality of first moments.

Abstract: Some aspects relate to integrated devices for obtaining timing and/or spectral information from incident light. In some embodiments, a pixel may include one or more charge storage regions configured to receive charge carriers generated responsive to incident photons from a light source, with charge carriers stored in the charge storage region(s) indicative of spectral and timing information. In some embodiments, a pixel may include regions having different depths, each configured to generate charge carriers responsive to incident photons. In some embodiments, a pixel may include multiple charge storage regions having different depths, and one or more of the charge storage regions may be configured to receive the incident photons and generate charge carriers therein. In some embodiments, a pixel may include an optical sorting element configured to direct at least some incident photons to one charge storage region and other incident photons to another charge storage region.

Abstract: A prediction algorithm determines synthetic fluorescence images on the basis of measurement images. A validation of the synthetic fluorescence images can be effected on the basis of reference images which are captured after the measurement images or are captured for a separate sample. Alternatively or additionally, a training of the prediction algorithm can be effected on the basis of training images which are captured after the measurement images or are captured for a separate sample.

Abstract: The invention relates to an apparatus 100 for 3D shaping of a workpiece 101 by material ablation with a laser beam 102. The apparatus 100 comprises a machining unit 103, which is configured to provide a pressurized fluid jet 104 onto the workpiece 101 and to couple the laser beam 102 into the fluid jet 104 towards the workpiece 101. Further, the apparatus 100 includes a motion controller 105 configured to set an x-y-z-position of the workpiece 101 relative to the machining unit 103. It also includes a measuring unit 107 configured to measure a z-position of the point of incidence 108 of the pressurized fluid jet 104 on the workpiece 101 in the z-direction.

Abstract: Described herein are techniques that improve the collection and readout of charge carriers in an integrated circuit. Some aspects of the present disclosure relate to integrated circuits having pixels with a plurality of charge storage regions. Some aspects of the present disclosure relate to integrated circuits configured to substantially simultaneously collect and read out charge carriers, at least in part. Some aspects of the present disclosure relate to integrated circuits having a plurality of pixels configured to transfer charge carriers between charge storage regions within each pixel substantially at the same time. Some aspects of the present disclosure relate to integrated circuits having three or more sequentially coupled charge storage regions. Some aspects of the present disclosure relate to integrated circuits capable of increased charge transfer rates.

Abstract: Systems, methods, and platforms for identifying and monitoring cell states are described. In one example, a computer-implemented method includes receiving cell imaging data comprising at least one transmitted light micrograph, generating a cell imaging dataset comprising time course data of transmitted light micrographs collected at an interval over a time, applying a machine learning model configured to analyze the cell imaging dataset to build a plurality of profiles of cell states, and identifying a cell state based on the plurality of profiles.

Abstract: Luminescent taggant compositions, luminescent materials that include luminescent taggants, and articles including luminescent taggants are provided herein. In an embodiment, a luminescent taggant composition includes a first luminescent taggant, a second luminescent taggant, and a third luminescent taggant. The first luminescent taggant includes a first emitting ion that produces a first emission in a first taggant emission band when exposed to excitation energy. The second luminescent taggant includes a second emitting ion that is different from the first emitting ion and that produces a second emission in a second taggant emission band that is different from the first taggant emission band when exposed to excitation energy. The first luminescent taggant is substantially free of the second emitting ion and the second luminescent taggant is substantially free of the first emitting ion. The third luminescent taggant includes the first emitting ion and the second emitting ion.

Abstract: A way to design a codebook for estimating the type of a molecule at a particular location in a fluorescence microscopy image makes use of one or both of (1) knowledge of the non-uniform prior distribution of molecule types (i.e., some types are known a priori to occur more frequently than others) and/or knowledge of co-occurrence of molecule types at close locations (e.g., in a same cell); and (2) knowledge of a model of the (e.g., random) process that yields the intensities that are expected at a location when a molecule with a particular subset of markers (i.e., a molecule of a type that has been assigned a codeword that defines that subset) is present at that location. The codebook design may provide experimental efficiency by reducing the number of images that need to be acquired and/or improve classification or detection accuracy by making the codewords for different molecule types more distinctive.

Abstract: A method for performing a surgical procedure includes planning a resection of a bone of a patient. A volume of the bone is removed according to the planned resection using a surgical tool. As the bone is removed, data corresponding to a shape and volume of the removed bone is tracked with a computer system operatively coupled to the surgical tool. A prosthesis is implanted onto the bone of the patient based on the tracked data corresponding to the shape of the removed bone.

Abstract: A microscope includes a wide-field illuminator configured to illuminate at least one selected region of a sample, and a beam splitter configured to generate a first detection beam path and a second detection beam path. A camera detector is arranged within the first detection beam path and is configured to record images of the selected region of the sample. A point detector is arranged within the second detection beam path and is configured to acquire a predetermined subregion of the sample lying within the selected region. A detection objective, which is arranged within the first and second detection beam paths on an object side of the beam splitter. The detection objective is a common detection objective for the camera detector and the point detector.

Abstract: Milling with ultraviolet excitation (MUVE) realizes high-throughput multiplex imaging of large three-dimensional samples. The instrumentation may comprise a UV-source attachment, precision stage attachment, and/or a blade assembly, and the instrumentation may overcome several constraints inherent to current state-of-the-art three-dimensional microscopy. MUVE offers throughput that is orders of magnitude faster than other technology by collecting a two-dimensional array of pixels simultaneously. The proposed instrumentation also utilizes serial ablation and provides the opportunity for true whole-organ imaging at microscopic resolution.

Abstract: A fluorescence assay device performs a fluorescence assay of one or more samples including fluorescent materials, causing a plurality of sets of datapoint to be stored relating to received fluorescence emitted by the fluorescent materials. A user interface displays representations of some of the sets of datapoints. A first region of the user interface includes alphanumeric representations of a subset of the plurality of sets of datapoints, including one or more user-specified sets of datapoints. A second region of the user interface includes one or more pictographic representations of the one or more user-selected sets of datapoints. The one or more pictographic representations indicate categorizations of values of the particular set of datapoints for the one or more samples.

Abstract: An inspection apparatus is an inspection apparatus for inspecting a sample on which a plurality of light-emitting elements is formed, and includes an excitation light source that generates excitation light to irradiate the sample, a camera that images fluorescence having a wavelength longer than a reference wavelength in fluorescence from the light-emitting elements, and a control apparatus that determines a quality of each of the light-emitting elements based on fluorescence imaged by the camera, in which the reference wavelength is a wavelength obtained by adding a full width at half maximum of a normal fluorescence spectrum of the light-emitting element to a peak wavelength of the normal fluorescence spectrum.

Abstract: Systems, methods, and devices for hyperspectral imaging in a light deficient environment are disclosed. A system includes an emitter for emitting pulses of electromagnetic radiation and an image sensor comprising a pixel array for sensing reflected electromagnetic radiation. The system includes a controller comprising a processor in electrical communication with the image sensor and the emitter, wherein the controller synchronizes timing of the pulses of electromagnetic radiation during a blanking period of the image sensor. The system is such that at least a portion of the pulses of electromagnetic radiation emitted by the emitter comprises one or more of electromagnetic radiation having a wavelength from about 513 nm to about 545 nm, electromagnetic radiation having a wavelength from about 565 nm to about 585 nm, or electromagnetic radiation having a wavelength from about 900 nm to about 1000 nm.

Abstract: Systems, methods, and instrumentalities are disclosed for data processing and creating a record of the processing for archival in metadata associated with the results of the processing. The processing may include transformations of the data. Transforming the data may generate transformed data. The processes performed may be archived, for example, in metadata associated with the transformed data. The metadata may be annotated with information associated with previous transforms performed on the transformed data. The metadata may be stored with the transformed data.

Abstract: Systems for detecting light (e.g., in a flow stream) are described. Light detection systems according to certain embodiments include a wavelength separator configured to generate first, second and third predetermined spectral ranges of light from a light source and first, second and third light detection modules configured to receive each of the first, second and third predetermined spectral ranges of light, the light detection modules having a plurality of photodetectors and an optical component that conveys light having a predetermined sub-spectral range to the photodetectors. Systems and methods for measuring light emitted by a sample (e.g., in a flow stream) and kits having three or more wavelength separators, a plurality of photodetectors and an optical component are also provided.

Abstract: A target may be tagged, and its position tracked to maintain the position of the target at the center (e.g., zero region) of a suppression beam, where suppression is minimal, so the signal from the target is not suppressed relative to signal from background molecules. Point scanning is not needed, as a pixelated detector of a camera can be used to acquire the position of the target in one shot. Images of the object from the pixelated detector can be analyzed to acquire the position of the target. To maintain the target in the zero, the beams may be steered onto the target. Alternatively or additionally, the sample may be moved to place the sample in the center. A correction signal may be sent to cause the sample and/or beams to be moved to maintain the target in position.

Abstract: An inspection apparatus includes a light source unit, cameras, a keyboard, and a controller that determines a wavelength of the excitation light, based on the information on the emission color received by the keyboard, and that controls the light source unit so that the light source unit generates excitation light with the determined wavelength. The controller determines a wavelength longer than an absorption edge wavelength of the substrate of the sample and shorter than a peak wavelength of an emission spectrum of the light-emitting element, the peak wavelength being specified from the information on the emission color, to be the wavelength of the excitation light.

Abstract: The present disclosure provides a method and apparatus for biomolecular multiplexed imaging through the iterative unmixing of fluorophores. According to the first embodiments of the present disclosure, although signals of two fluorophores are detected in the first fluorescent detection spectral range, the signals of the two images obtained from two detection spectral ranges can be unmixed through the iterative minimization of mutual information. Furthermore, the present disclosure provides a multi-color unmixing method and apparatus through the iterative minimization of mutual information. In the second embodiments of the present disclosure, a plurality of images of a plurality of fluorophores marking different biomolecules, respectively, for example, N fluorophores are obtained. Images, each containing the signals of single fluorophore can be obtained from the obtained images while minimizing the mutual information shared by images of each of pairs each consisting of two of the obtained images.

Abstract: Optical traps enable nanoscale manipulation of individual biomolecules while measuring molecular forces and lengths. Disclosed herein is a camera-based detection system that enables accurate and precise measurements of forces and interactions in a dual optical trap. Optical traps may be used to stretch and unzip DNA molecules while measuring the displacements of trapped particles from their trapping centers with sub-nanometer accuracy and precision.

Abstract: System and methods for analyzing single molecules and performing nucleic acid sequencing. An integrated device includes multiple pixels with sample wells configured to receive a sample, which when excited, emits radiation. The integrated device includes at least one waveguide configured to propagate excitation energy to the sample wells from a region of the integrated device configured to couple with an excitation energy source. A pixel may also include at least one element for directing the emission energy towards a sensor within the pixel. The system also includes an instrument that interfaces with the integrated device. The instrument may include an excitation energy source for providing excitation energy to the integrated device by coupling to an excitation energy coupling region of the integrated device.

Abstract: A plastic item, such as a beverage bottle, conveys two distinct digital watermarks, encoded using two distinct signaling protocols. A first, printed label watermark conveys a retailing payload, including a Global Trade Item Number (GTIN) used by a point-of-sale scanner in a retail store to identify and price the item when presented for checkout. A second, plastic texture watermark conveys a recycling payload, including data identifying the composition of the plastic. The use of two different signaling protocols assures that a point-of-sale scanner will not spend its limited time and computational resources working to decode the recycling watermark, which lacks the data needed for retail checkout. In some embodiments, a recycling apparatus makes advantageous use of both types of watermarks to identify the plastic composition of the item (e.g., relating GTIN to plastic type using an associated database), thereby increasing the fraction of items that are correctly identified for sorting and recycling.

Abstract: Devices and methods for visibly highlighting areas of a region including an imager configured to image the region with a sensitivity to at least one of wavelength, light level, or contrast greater than the human eye, an overlay element configured to visibly highlight areas of the region and registered to the imager to produce alignment of imaged features with highlighted features at the same location on the region, and at least one of a controller executing a program or logic configured to process acquired images from the imager to identify areas of the region determined not visible to the human eye, and control the overlay element to visibly highlight those areas on the region.

Abstract: A method of evaluating quality of graphene using a confocal laser scanning microscope, the method comprising steps of: irradiating with laser light a graphene layer formed on a catalyst layer; detecting a signal of light reflected from the graphene layer; forming a planar image of the graphene layer using the detected optical signal; and analyzing a contrast between graphene and the catalyst layer in the planar image, wherein the graphene layer is continuously formed on the catalyst layer by a roll-to-roll process.

Abstract: An apparatus for determining a characteristic of a photoluminescent (PL) layer comprises: a light source that generates an excitation light that includes light from the visible or near-visible spectrum; an optical assembly configured to direct the excitation light onto a PL layer; a detector that is configured to receive a PL emission generated by the PL layer in response to the excitation light interacting with the PL layer and generate a signal based on the PL emission; and a computing device coupled to the detector and configured to receive the signal from the detector and determine a characteristic of the PL layer based on the signal.

Abstract: Detecting emission light, in a laser scanning microscope, wherein the emission light emanating from a sample is guided onto a two-dimensional matrix sensor having a plurality of pixels and being located on an image plane, and a detection point distribution function is detected by the matrix sensor in a spatially oversam pled manner. The emission light emanating from the sample is spectrally separated in a dispersion device; the spectrally separated emission light is detected by the matrix sensor in a spectrally resolved manner; and during the analysis of the intensities measured by the pixels of a pixel region, the spectral separation is cancelled at least for some of said pixels. Additional aspects relate to a detection device for the spectrally resolved detection of emission light in a laser scanning microscope and to a laser scanning microscope.

Abstract: The present invention is directed toward a system and method for STED nanography, which reduces background noise. To remove background noise from a STED image, the polarization of the STED beam is altered from that used to obtain the original image. A polarized image is obtained. This polarized image can then be subtracted from the original image to remove noise inherent to the image.

Abstract: The present disclosure provides a system and method for the detection of rare mutations and copy number variations in cell free polynucleotides. Generally, the systems and methods comprise sample preparation, or the extraction and isolation of cell free polynucleotide sequences from a bodily fluid; subsequent sequencing of cell free polynucleotides by techniques known in the art; and application of bioinformatics tools to detect rare mutations and copy number variations as compared to a reference. The systems and methods also may contain a database or collection of different rare mutations or copy number variation profiles of different diseases, to be used as additional references in aiding detection of rare mutations, copy number variation profiling or general genetic profiling of a disease.

Abstract: A two-photon stimulated emission depletion composite microscope using continuous light loss, the microscope comprising: a two-photon imaging unit (100) and an STED imaging unit (200), wherein for thicker samples, the two-photon imaging unit (100) can be used, and for regions of interest on the surface of samples, the STED super-resolution imaging unit (200) can be used. The two-photon stimulated emission depletion composite microscope using continuous light loss integrates two functions of STED imaging and two-photon imaging, so as to provide a powerful tool for cutting-edge biomedical research.

Abstract: Fluorescence microscopy method comprising: illuminating a scattering sample with a coherent excitation light beam having a wavefront with an initial configuration for exciting fluorescent emitters in the scattering sample; acquiring an initial image; selecting target pixels in an area of the initial image; optimising the initial configuration of the wavefront for each target pixel for decreasing the speckle grain size and obtaining a final image; subtracting the initial image from the final image for each target pixel, obtaining an image; fitting the image with a Gaussian function with free center coordinates for each target pixel; generating an image containing a Gaussian distribution centered at the coordinates and intensity, and with a waist equal to an average size S of speckle grain of the scattering sample, for each target pixel; and generating a final image of the sample by summing the images of the target pixels.

Abstract: An apparatus for analysis of a sample of a material is disclosed. The apparatus has a body configured to accept the sample and a detector with a plurality of points. The detector is configured to provide a signal about the intensities of light received at the points. The apparatus also includes a slit array having a plurality of slits configured to collected light from the accepted sample and a collimating lens array having a plurality of lenses configured to receive the light from the slit array and collimate the collected light. The apparatus also includes a diffraction grating configured to diffract the light received from the collimating lens array and a focusing lens configured to focus the diffracted light of a common frequency on a common point of the detector.

Abstract: Techniques are disclosed for detecting a presence of a biological substance through an article such as a diaper. For example, a detection system causes a light source to transmit light through the article. The light includes a peak wavelength that corresponds to an excitation wavelength of a biological substance that may be present in the article. The detection system detects a measurement of light intensity within a range of emission wavelengths of the biological substance. By comparing the measurement of light intensity to a threshold, the detection system identifies a presence of the biological substance on the article.

Abstract: A method for performing a surgical procedure includes planning a resection of a bone of a patient. A volume of the bone is removed according to the planned resection using a surgical tool. As the bone is removed, data corresponding to a shape and volume of the removed bone is tracked with a computer system operatively coupled to the surgical tool. A prosthesis is implanted onto the bone of the patient based on the tracked data corresponding to the shape of the removed bone.

Abstract: Systems for detecting light (e.g., in a flow stream) are described. Light detection systems according to certain embodiments include a wavelength separator configured to generate first, second and third predetermined spectral ranges of light from a light source and first, second and third light detection modules configured to receive each of the first, second and third predetermined spectral ranges of light, the light detection modules having a plurality of photodetectors and an optical component that conveys light having a predetermined sub-spectral range to the photodetectors. Systems and methods for measuring light emitted by a sample (e.g., in a flow stream) and kits having three or more wavelength separators, a plurality of photodetectors and an optical component are also provided.

Abstract: The present disclosure provides an experiment platform for simulating fire in an underground traffic conversion channel, including: a model body configured to simulate fire in the an underground traffic conversion channel; a burner connected to the model body and configured to generate smoke; and a smoke imaging system including laser sheet light sources and image recording devices configured to record smoke distribution images. The laser sheet light sources are in the model body, and plane laser light emitted by the laser sheet light sources is parallel to a flow direction of the smoke. The image recording devices are in one-to-one correspondence to the laser sheet light sources and are arranged outside an observation window of the model body. One laser sheet light source and one image recording device corresponding thereto each have a filter configured to filter out laser light of a same wavelength.

Abstract: The present disclosure provides a detection device capable of detecting a low concentration of an analyte with high sensitivity. The detection apparatus according to the present disclosure comprises a metal microstructure on which a first VHH antibody having a property of binding specifically to the analyte is immobilized and which generate surface plasmon by being irradiated with excitation light, an inlet through which a second VHH antibody and a sample that may contain an analyte are introduced, wherein the second VHH antibody has a property of binding specifically to the analyte and is labeled with a fluorescent substance, a light source for irradiating the metal microstructure to which the second VHH antibody and the sample have been introduced with the excitation light, and a detection unit for detecting the analyte on the basis of fluorescence generated from the fluorescent substance by the irradiation of the excitation light.

Abstract: A low-powered sensor, a method for a low-powered sensor, and an apparatus are provided for monitoring a condition of a machine. Responsive to receiving a trigger signal during a setup phase, the low-powered sensor collects sensor data for a predetermined extended period of time. The low-powered sensor receives a setting of a delay based on a determined amount of time of the predetermined extended period of time, since the receiving of the trigger signal, for the machine to reach a steady operating condition within predetermined limits. Upon receiving the trigger signal after the receiving of the setting of the delay, the low-powered sensor waits a period of time of the set delay before collecting the sensor data for a predetermined amount of time that is shorter than the predetermined extended period of time, and provides the collected sensor data to a computing device for analysis.

Abstract: The invention provides systems and methods for imaging a sample. In various embodiments, the invention provides a system comprising an image sensor, a laser for emitting excitation light for an infrared or near-infrared fluorophore, a visible light source, a notch beam splitter, a notch filter, a synchronization module, an image processing unit, an image displaying unit, and light-conducting channels. In various embodiments, the present invention provides a system comprising an image sensor, a laser for emitting excitation light for an infrared or near-infrared fluorophore, a laser clean-up filter, a notch filter, a white light source, an image processing unit, an image displaying unit, and light-conducting channels. In accordance with the present invention, the image sensor can detect both visible light and infrared light.

Abstract: Methods for characterizing spillover spreading originating from a first fluorochrome in fluorescent flow cytometer data collected for a second fluorochrome are provided. In some embodiments, methods include partitioning the fluorescent flow cytometer data according to the intensity of the data relative to the first fluorochrome. In embodiments, methods also include estimating with a first linear regression a zero-adjusted standard deviation for the intensity of light collected from the second fluorochrome for each of the partitioned quantiles based on the assumption that the intensity of light collected from the first fluorochrome is zero, and obtaining with a second linear regression a spillover spreading coefficient from the zero-adjusted standard deviations. Systems and computer-readable media for characterizing spillover spreading originating from a first fluorochrome in fluorescent flow cytometer data collected for a second fluorochrome are also provided.

Abstract: An apparatus for determining a characteristic of a photoluminescent (PL) layer comprises: a light source that generates an excitation light that includes light from the visible or near-visible spectrum; an optical assembly configured to direct the excitation light onto a PL layer; a detector that is configured to receive a PL emission generated by the PL layer in response to the excitation light interacting with the PL layer and generate a signal based on the PL emission; and a computing device coupled to the detector and configured to receive the signal from the detector and determine a characteristic of the PL layer based on the signal.

Abstract: A light detection sensor includes a light source that irradiates a detection target with light, a rod lens that collects emitted light from the detection target, a first light receiving element that detects the emitted light that has passed through the rod lens, and a second light receiving element that detects the emitted light that has passed through the rod lens, at a position different from a position at which the first light receiving element detects the light.

Abstract: A means to enable the inspection of industrial materials in order to see hidden structural defects that can lead to an early failure, using electromagnetic fields and harmonic waves.

Abstract: A fluorescence scanning microscope includes excitation and de-excitation light sources, which are designed to generate an excitation and a de-excitation light distribution, respectively. An illumination unit combines the light distributions to form a light distribution scanning over multiple illumination target points of a sample in such a way that an intensity maximum of the excitation light distribution and an intensity minimum of the de-excitation light distribution are spatially superimposed on one another. A detector detects fluorescence photons emitted from the respective illumination target point as a function of their arrival times. A processor evaluates the fluorescence photons with respect to the arrival times, generates a first pixel and a second pixel based thereon, assembles the first and second pixels to form first and second sample images, respectively, and, by means of the two sample images, determines a spatial offset between the intensity maximum and the intensity minimum.

Abstract: The invention relates to a photoplethysmography (PPG) sensing device comprising —a pulsed light source, —at least one pixel to create photo-generated electrons, synchronized with said pulsed light source. It is mainly characterized in that each pixel comprises: —a pinned photodiode (PPD) having two electronic connection nodes, —a sense node (SN), to convert the photo-generated electrons into a voltage, and —a Transfer Gate (TGtransfer) transistor, having its source electronically connected to one electronic connection node of said pinned photodiode (PPD), and being configured to act as a transfer gate (TG) between said pinned photodiode (PPD) and said sense node (SN), allowing the photo-generated electrons to sink when the light is pulsed-off, the photo-generated electrons integration when the light is pulsed-on and the transfer of at least part of the integrated photo-generated electrons to said sense node for a read-out.

Abstract: A two-photon stimulated emission depletion composite microscope, comprising a two-photon imaging unit (100) and an STED imaging unit (200), wherein the two-photon imaging unit (100) can be used for a relatively thick sample, and the STED super-resolution imaging unit can be used for a region of interest on a surface of a sample, and the microscope makes light spots generated by an excitation light and a depletion light after being focused by an objective lens (OL) accurately coincide in a three-dimensional distribution. The two-photon stimulated emission depletion composite microscope (10) integrates two functions of STED imaging and two-photon imaging and makes the two types of light spots generated by an excitation light and a depletion light after being focused by an objective lens accurately coincide in a three-dimensional distribution, thereby providing a powerful tool for cutting-edge biomedical research.

Abstract: Aspects of the present disclosure include methods for spectrally resolving light from fluorophores having overlapping fluorescence spectra in a sample. Methods according to certain embodiments include detecting light with a light detection system from a sample having a plurality of fluorophores having overlapping fluorescence spectra and spectrally resolving light from each fluorophore in the sample. In some embodiments, methods include estimating the abundance of one or more of the fluorophores in the sample, such as on a particle. In certain instances, methods include identifying the particle in the sample based on the abundance of each fluorophore and sorting the particle. Methods according to some embodiments includes spectrally resolving the light from each fluorophore by calculating a spectral unmixing matrix for the fluorescence spectra of each fluorophore. Systems and integrated circuit devices (e.g., a field programmable gate array) for practicing the subject methods are also provided.

Abstract: An observation device is an observation device observing an observation target and includes: an emission light source that generates emission light; a projection light source that generates projection light; a scanning mirror that scans the emission light and the projection light toward the observation target along the same optical path; a light guide optical system that guides detection target light generated in the observation target in accordance with emission of the emission light without it passing through the scanning mirror; an optical detector that detects the detection target light guided by the light guide optical system; and a control unit that controls an intensity of the projection light on the basis of a result of the detection of the detection target light.

Abstract: Methods for classifying fluorescent flow cytometer data are provided. In some instances, methods include processing the flow cytometer data with a supervised algorithm configured to cluster the fluorescent flow cytometer data into distinct populations according to the relationship of data points to relevant threshold values. In embodiments, methods include determining a measure of spillover spreading by calculating spillover spreading coefficients and combining them in a spillover spreading matrix. In some embodiments, populations of fluorescent flow cytometer data are adjusted to subtract the magnitude of spillover spreading. In embodiments, spillover spreading adjusted populations are partitioned after potential partitions are evaluated relative to the threshold values. In embodiments, partitioned populations of fluorescent flow cytometer data are classified (i.e., phenotyped) according to a hierarchy. Systems and computer-readable media for classifying fluorescent flow cytometer data are also provided.