Aspects of the present disclosure include methods for modulating an intensity profile of a laser beam. Methods according to certain embodiments include irradiating an acousto-optic device with a laser to generate an output laser beam having a plurality of angularly deflected laser beams, capturing an image of the output laser beam, determining an intensity profile of the output laser beam along a horizontal axis from the captured image and adjusting one or more parameters of a waveform inputted into the acousto-optic device in response to the determined intensity profile to generate an output laser beam having a modulated intensity profile. Systems having a laser, an acousto-optic device, an imaging sensor and a waveform generator as well as non-transitory computer readable storage medium with instructions for practicing the subject methods are also described.

An imaging flow cytometer includes: a laser unit that emits first and second laser light to first and second spots; a first and second imaging sections that image the first and second spots; first and second detection devices that detect a particle that passes through the first and second spots; a first particle detection section that issues an imaging timing instruction signal to the first and second imaging sections; an image storage section that receives an image imaged by the first and second imaging sections; and a sorting determination section that determines whether the particle is an objective particle. The first and second imaging sections clip an image of the particle based on the imaging timing instruction signal.

Described herein are apparatuses for analyzing an optical signal decay. In some embodiments, an apparatus includes: a source of a beam of pulsed optical energy; a sample holder configured to expose a sample to the beam; a detector comprising a number of spectral detection channels configured to convert the optical signals into respective electrical signals; and a signal processing module configured to perform a method. In some embodiments, the method includes: receiving the electrical signals from the detector; mathematically combining individual decay curves in the electrical signals into a decay supercurve, the supercurve comprising a number of components, each component having a time constant and a relative contribution to the supercurve; and numerically fitting a model to the supercurve.

A fractionated photoacoustic flow cytometry (PAFC) system and methods for the in vivo detection of target objects in biofluidic systems (e.g., blood, lymph, urine, or cerebrospinal fluid) of a living organism is described. The fractionated system includes a fractionated laser system, a fractionated optical system, a fractionated acoustic system, and combinations thereof. The fractionated laser system includes at least one laser or laser array for pulsing a target object within the circulatory vessel with fractionated focused laser beams. The fractionated optical system separates one or several laser beams into multiple beams in a spatial configuration on the skin above the circulatory vessel of the living organism. The fractionated acoustic system includes multiple focused ultrasound transducers for receiving photoacoustic signals emitted by the target object in response to the fractionated laser beams. The target objects have intrinsic photoacoustic contrast or may be labeled with photoswitchable or spaser-based probes. Fractioned beams may be used also for diagnostics with other spectroscopic methods (e.g., fluorescence, Raman or scattering) and energy sources both coherent and conventional such as lamp and LED in the broad spectral range from 10 Å to 1 cm (e.g., X-ray, UV, visible, NIR or microwaves) in continuous wave and pulse modes.

A kit and method for flow cytometry include a liquid dye concentrate for fluorescent staining of virus-size particles with a plurality of fluorogenic dyes in a liquid medium. The liquid dye concentrate includes a plurality of fluorogenic dyes and one or both of (i) the liquid medium comprising a liquid mixture including water and liquid phase organic material and (ii) disaccharide dissolved in the liquid medium.

Aspects of the present disclosure include systems for irradiating particles in a flow stream. Systems according to certain embodiments include a light source having a first laser configured for continuous irradiation of a flow stream and a second laser configured for irradiation of the flow stream in discrete intervals where each discrete interval of irradiation by the second laser is triggered by irradiation of a particle in the flow stream with the first laser. Methods for irradiating a sample in a flow stream with the subject light sources are also described. Computer readable storage medium for practicing the subject methods are provided. Kits having one or more lasers are also provided.

The present disclosure provides apparatuses, systems, and methods for performing particle analysis through flow cytometry at comparatively high event rates and for gathering high resolution images of particles.

The present disclosure provides apparatuses, systems, and methods for performing particle analysis through flow cytometry at comparatively high event rates and for gathering high resolution images of particles.

Disclosed is an apparatus and method for fluorescence excitation and detection. The apparatus comprises one or more light sources for providing excitation light for fluorescence excitation at an observation spot along an optical axis for excitation, an optical collection element for collecting fluorescence light generated by the excitation light at two or more different observation spots into two or more different measurement channels with an optical axis for collecting non-parallel to the optical axis for excitation of each of the one or more light sources, and, for each of the two or more measurement channels and thereby for each of the two or more observation spots, a dedicated optical detector for detecting fluorescence from the fluorescence light collected by the optical collection element.

A crystal for flow cytometry with dual laser beams is disclosed. The crystal is a birefringent crystal comprising a material composition including a quartz mineral having a face side including a face angle of ninety degrees plus or minus one tenth of a degree; a wedge side that is substantially perpendicular to the face side, wherein the wedge side includes a wedge angle of two degrees plus or minus one tenth of a degree; and a major side that is substantially perpendicular to the face side and the wedge side. The major side includes a thickness of one and one-half millimeter plus or minus one tenth of a millimeter. A polarized light beam entering the birefringent crystal at an incident angle is separated into an ordinary light beam and an extraordinary light beam.