Abstract: Systems having an unfiltered light scatter detector configured to detect scattered light from a sample in a flow stream are provided. Systems according to certain embodiments include a light source having two or more lasers, a light detection system having an unfiltered light scatter detector and a processor having memory operably coupled to the processor where the memory includes instructions which when executed by the processor, cause the processor to generate one or more data signals in response to scattered light from each of the two or more lasers detected by the unfiltered light scatter detector; and determine one or more parameters of data acquisition based on the generated data signals from the unfiltered light scatter detector. Methods for determining one or more parameters for data acquisition with the subject systems are also described.

Abstract: Aspects of the present disclosure include methods for adjusting sensitivity of a photodiode in a light detection system. Methods according to certain embodiments include detecting light with a light detection system having a photodiode and an amplifier, determining responsivity of the photodiode over a plurality of wavelengths of light and adjusting one or more parameters of the amplifier in response to the responsivity of the photodiode over the plurality of wavelengths of light. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a photodiode and an amplifier for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.

Abstract: Flow cytometers having sample injection needles are provided. The subject flow cytometers include a flow cell for transporting particles in a core stream of a flow stream from a proximal end to a distal end, and a sample injection needle having a passage running therethrough for delivering sample fluid from a sample injection line at a proximal end to the flow cell at a distal end to generate the core stream. Flow cells of interest include a flow cell cone at the proximal end, and sample injection needles of interest are configured and positioned to maintain an intact core stream under flow conditions that vary by a magnitude or more. Methods of assembling the subject flow cytometers and methods of analyzing a sample using the subject flow cytometers are also provided.

Abstract: Aspects of the present disclosure include methods for adjusting sensitivity of a photodiode in a light detection system. Methods according to certain embodiments include detecting light with a light detection system having a photodiode and an amplifier, determining responsivity of the photodiode over a plurality of wavelengths of light and adjusting one or more parameters of the amplifier in response to the responsivity of the photodiode over the plurality of wavelengths of light. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a photodiode and an amplifier for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.

Abstract: Aspects of the present disclosure include methods for determining absolute count of particles in a sample in a flow cytometer. Methods according to certain embodiments include introducing a bubble into a flow stream propagating a sample having particles, irradiating the flow stream with a light source, detecting light from the irradiated particles with a photodetector, detecting the presence of the bubble in the flow stream with the photodetector and determining the absolute count of the particles in the sample based on data signals generated in response to light detected from the irradiated particles, a data signal generated when the bubble is introduced into the flow stream and data signals generated in response to the detected bubble. Systems (e.g., flow cytometers) having a light source, a light detection system and a sample line that is configured to introduce a bubble into a flow stream for practicing the subject methods are also described.

Abstract: Methods for evaluating flow cytometer data for the presence of a coincident event are provided. Methods of interest include receiving flow cytometer data associated with a first particle, where the received flow cytometer data is obtained from a detector following the irradiation of the first particle and a second particle in a flow stream by first and second light sources, respectively. Methods also include receiving first and second trigger signals associated with the first and second particles, respectively. In addition, the subject methods include generating a time interval for the first and second particles based on a difference between the first and second trigger signals, and evaluating the flow cytometer data for the presence of a coincident event based on the generated time interval by determining if the first and second particles are irradiated by the first and second light sources simultaneously. Systems and computer-readable media are also provided.

Abstract: 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.

Abstract: Systems having an unfiltered light scatter detector configured to detect scattered light from a sample in a flow stream are provided. Systems according to certain embodiments include a light source having two or more lasers, a light detection system having an unfiltered light scatter detector and a processor having memory operably coupled to the processor where the memory includes instructions which when executed by the processor, cause the processor to generate one or more data signals in response to scattered light from each of the two or more lasers detected by the unfiltered light scatter detector; and determine one or more parameters of data acquisition based on the generated data signals from the unfiltered light scatter detector. Methods for determining one or more parameters for data acquisition with the subject systems are also described.

Abstract: Methods and systems for determining a drop delay of a flow stream in a flow cytometer are provided. Aspects of the methods according to certain embodiments include capturing an image of the flow stream to obtain an imaged flow stream, identifying a disturbance at a break off point in the imaged flow stream and calculating the drop delay of the flow stream based on the identified disturbance. Systems for practicing the subject methods having an imaging sensor for capturing one or more images of the flow stream and a processor configured to identify a disturbance at a break off in the imaged flow stream and calculating the drop delay using the imaged flow stream are also provided. Non-transitory computer readable storage mediums are also described.

Abstract: Aspects of the present disclosure include a particle sorting module for sorting components of a sample, such as cells in a biological sample. Particle sorting modules according to certain embodiments include an enclosed housing having an aligner for coupling the housing with a particle sorting system, a flow cell nozzle positioned at the proximal end of the housing, a sample interrogation region in fluid communication with the orifice of the flow cell nozzle and a droplet deflector. A particle sorting system and methods for separating components of a sample as well as kits, including one or more particle sorting module, suitable for coupling with a particle sorting system and for practicing the subject methods are also provided.

Abstract: Systems having an unfiltered light scatter detector configured to detect scattered light from a sample in a flow stream are provided. Systems according to certain embodiments include a light source having two or more lasers, a light detection system having an unfiltered light scatter detector and a processor having memory operably coupled to the processor where the memory includes instructions which when executed by the processor, cause the processor to generate one or more data signals in response to scattered light from each of the two or more lasers detected by the unfiltered light scatter detector; and determine one or more parameters of data acquisition based on the generated data signals from the unfiltered light scatter detector. Methods for determining one or more parameters for data acquisition with the subject systems are also described.

Abstract: Aspects of the present disclosure include methods for adjusting sensitivity of a photodiode in a light detection system. Methods according to certain embodiments include detecting light with a light detection system having a photodiode and an amplifier, determining responsivity of the photodiode over a plurality of wavelengths of light and adjusting one or more parameters of the amplifier in response to the responsivity of the photodiode over the plurality of wavelengths of light. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a photodiode and an amplifier for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.

Abstract: Systems having an unfiltered light scatter detector configured to detect scattered light from a sample in a flow stream are provided. Systems according to certain embodiments include a light source having two or more lasers, a light detection system having an unfiltered light scatter detector and a processor having memory operably coupled to the processor where the memory includes instructions which when executed by the processor, cause the processor to generate one or more data signals in response to scattered light from each of the two or more lasers detected by the unfiltered light scatter detector; and determine one or more parameters of data acquisition based on the generated data signals from the unfiltered light scatter detector. Methods for determining one or more parameters for data acquisition with the subject systems are also described.

Abstract: 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.

Abstract: Apparatus and methods are described for automatically performing set-up steps for flow cytometry operations. The invention provides for the spatial determination of a flow stream and the subsequent automatic alignment of analysis devices and/or collection vessels. The automatic determination of flow stream properties provides for the automatic configuration flow cytometer parameters.

Abstract: Methods and systems for determining a drop delay of a flow stream in a flow cytometer are provided. Aspects of the methods according to certain embodiments include capturing an image of the flow stream to obtain an imaged flow stream, identifying a disturbance at a break off point in the imaged flow stream and calculating the drop delay of the flow stream based on the identified disturbance. Systems for practicing the subject methods having an imaging sensor for capturing one or more images of the flow stream and a processor configured to identify a disturbance at a break off in the imaged flow stream and calculating the drop delay using the imaged flow stream are also provided. Non-transitory computer readable storage mediums are also described.

Abstract: Methods and systems for determining a drop delay of a flow stream in a flow cytometer are provided. Aspects of the methods according to certain embodiments include capturing an image of the flow stream to obtain an imaged flow stream, identifying a disturbance at a break off point in the imaged flow stream and calculating the drop delay of the flow stream based on the identified disturbance. Systems for practicing the subject methods having an imaging sensor for capturing one or more images of the flow stream and a processor configured to identify a disturbance at a break off in the imaged flow stream and calculating the drop delay using the imaged flow stream are also provided. Non-transitory computer readable storage mediums are also described.

Abstract: Apparatus and methods are described for automatically performing set-up steps for flow cytometry operations. The invention provides for the spatial determination of a flow stream and the subsequent automatic alignment of analysis devices and/or collection vessels. The automatic determination of flow stream properties provides for the automatic configuration flow cytometer parameters.

Abstract: Apparatus and methods are described for automatically performing set-up steps for flow cytometry operations. The invention provides for the spatial determination of a flow stream and the subsequent automatic alignment of analysis devices and/or collection vessels. The automatic determination of flow stream properties provides for the automatic configuration flow cytometer parameters.