A flow cytometer can include: at least one light emitter configured to emit light in a light path; a rectangular flow cell having flow cell width that is substantially lateral to the light path and a flow cell depth that is longitudinal to the light path, wherein the light path has an interrogation width at the flow cell that is narrower than the flow cell width; and a spherical reflector positioned adjacent to the rectangular flow cell and having a concave reflective surface that has a reflective direction that is positioned substantially orthogonal with the light path such that reflected light is reflected along a reflected path that is substantially orthogonal with the light path. At least one light absorbing member is positioned at least partially around the reflected path to absorb reflected light at an angle to the reflected path.

Provided is a fluorescence reader that uses two excitation channels and can read up to seven different fluorescent dyes in a single run. Each excitation channel has one light source and one single excitation filter and one dichroic mirror. One excitation channel is capable of exciting multiple fluorescent dyes and can be used to distinguish multiple dyes in combination with multiple emission filters. The excitation channels are driven by a motor that can automatically switch the two excitation channels for taking images of up to seven different fluorescent dyes. An algorithm to calibrate the crosstalk between different fluorescent dyes is also provided. Also provided is a method for analyzing digital PCR data using a ratio of two fluorescence emission readings.

A reaction processing apparatus includes: a reaction processing vessel; a first fluorescence detection device that irradiates a sample with first excitation light and detects first fluorescence produced from the sample; and a second fluorescence detection device that irradiates a sample with second excitation light and detects second fluorescence produced from the sample. The wavelength range of the first fluorescence and the wavelength range of the second excitation light overlap at least partially. The first excitation light and the second excitation light flash at a predetermined duty ratio d. The phase difference between the flashing of the first excitation light and the flashing of the second excitation light is set within a range of 2π(pm−Δpm) (rad) to 2π(pm+Δpm) (rad) or within a range of 2π[(1−pm)−Δpm] (rad) to 2π[(1−pm)+Δpm] (rad), where pm=d−d2 and Δpm =0.01*pm.

A method for evaluating a biological material for unassociated virus-like particles virus size having a particular epitope uses a fluorescent antibody stain specific for binding with the epitope and a fluid sample with the virus-size particles and fluorescent antibody stain is subjected to flow cytometry with identification of fluorescent emission detection events indicative of passage through a flow cell of a flow cytometer of unassociated labeled particles of virus size including such a virus-like particle and fluorescent antibody stain.

A multicolor fluorescence analysis device 11 is for detecting fluorescence emitted, as a result of excitation light irradiation, from a plurality of types of fluorophores included in a sample s, and is provided with an irradiation optical unit 520 for irradiating light emitted from a light source 510 onto a sample s as excitation light, a fluorescence condensation unit 530 having a fluorescence filter 531 that transmits light emitted from the sample s and transmits light of transmission wavelength bands different from the excitation wavelength bands, and a two-dimensional detector 554 that has a plurality of types of transmission filters 556 for transmitting prescribed wavelengths of light and detects the intensity of the light of the prescribed wavelength for each transmission filter 556, and the light emitted from at least two fluorophores from among the plurality of types of fluorophores is detected simultaneously and the fluorophore types are identified accordingly.

A structured substrate includes a substrate body having an active side. The substrate body includes reaction cavities that open along the active side and interstitial regions that separate the reaction cavities. The structured substrate includes an ensemble amplifier positioned within each of the reaction cavities. The ensemble amplifier includes a plurality of nanostructures configured to at least one of amplify electromagnetic energy that propagates into the corresponding reaction cavity or amplify electromagnetic energy that is generated within the corresponding reaction cavity.

The present disclosure provides a real-time thermocycler, comprising: a well for storing a sample comprising a target and fluorescence molecules, a thermal unit for adjusting a temperature of the sample, an excitation unit for exciting the fluorescence molecules of the sample via radiation, a detection unit for detecting a fluorescence signal from the sample, and a controller for controlling the excitation unit to adjust an intensity of the excitation of the sample based on information about the target, such that the fluorescence signal is in a working range of the detection unit.

A microfluidic reaction chamber with a reaction chamber circuit includes a microfluidic reaction chamber to contain a reaction fluid for amplification of nucleic acids, and a reaction chamber circuit disposed within the microfluidic reaction chamber. The microfluidic reaction chamber includes a base wall, a top wall parallel to the base wall and defined in part by a transparent lid, a first side wall, and a second side wall. The reaction chamber circuit is disposed within the microfluidic reaction chamber, and includes a top surface, a bottom surface, a first side wall, and a second side wall. The reaction chamber circuit is in fluidic contact with the reaction fluid and includes a photodetector to detect a fluorescence signal from a labeled fluorescent tag in the reaction fluid.

A detection apparatus having a read head including a plurality of microfluorometers positioned to simultaneously acquire a plurality of the wide-field images in a common plane; and (b) a translation stage configured to move the read head along a substrate that is in the common plane. The substrate can be a flow cell that is included in a cartridge, the cartridge also including a housing for (i) a sample reservoir; (ii) a fluidic line between the sample reservoir and the flow cell; (iii) several reagent reservoirs in fluid communication with the flow cell, (iv) at least one valve configured to mediate fluid communication between the reservoirs and the flow cell; and (v) at least one pressure source configured to move liquids from the reservoirs to the flow cell. The detection apparatus and cartridge can be used together or independent of each other.

Compact optical sources and methods for producing short and ultrashort optical pulses are described. A semiconductor laser or LED may be driven with a bipolar waveform to generate optical pulses with FWHM durations as short as approximately 85 ps having suppressed tail emission. The pulsed optical sources may be used for fluorescent lifetime analysis of biological samples and time-of-flight imaging, among other applications.