Abstract: The disclosure provides example rinse station apparatus, cartridges, and methods for flow cytometry. The rinse station apparatus includes: (a) a cartridge docking station having a base with a recessed receptacle for a cartridge, a vertical support coupled to the base's first end and a top support coupled to the vertical support and cantilevered over the base, the top support has an opening that aligns with the cartridge's opening, (b) a locking arm coupled to the base's second end, the locking arm's free end has a ridge to cooperate with a detent coupled to the cartridge's rear wall to retain the cartridge in place, (c) a spring coupled to the vertical support's front face to apply force to the cartridge's front wall to bias the cartridge toward the locking arm, and (d) a load cell coupled to the base of the cartridge docking station, the load cell measure's the cartridge's weight.

Abstract: Systems, methods, and apparatuses of controlling fluid flow are disclosed. An apparatus includes a first microplate having a first open portion and defining one or more first wells therein, a second microplate having a second open portion and defining one or more second wells therein, and a pneumatic lid constructed of styrene ethylene butylene styrene (SEBS). The pneumatic lid extends over the first open portion and the second open portion and includes one or more microfluidic channels that fluidly couple the one or more first wells to the one or more second wells. The pneumatic lid provides an airtight seal over the first microplate and the second microplate.

Abstract: A flow cytometer module configured to be integrated with a liquid handling system is provided herein. The flow cytometer module includes (a) a flow cell, (b) a first fluidic pathway, (c) an inlet configured to receive a sample introduction device of the liquid handling system including one or more samples, (d) a second fluidic pathway in fluid communication with the first fluidic pathway, (e) a laser interrogation device configured to examine the one or more samples at a laser interrogation point in the second fluidic pathway, and (f) a controller in communication with the liquid handling system and configured to cause the flow cytometer module to perform functions comprising: (i) recording data from the laser interrogation device corresponding to a plurality of events as the one or more samples pass the laser interrogation point, and (ii) transmitting the data corresponding to the plurality of events to the liquid handling system.

Abstract: A flow cytometer apparatus is provided herein including (a) a flow cell, (b) a fluidic pathway having a second end coupled to a first end of the flow cell, (c) a probe coupled to a first end of the fluidic pathway, (d) a sensor configured to detect one or more properties of a fluid in the fluidic pathway and positioned between the probe and the first end of the flow cell, (e) a processor in communication with the sensor, and (f) a non-transitory computer readable medium having stored therein instructions that are executable to cause the processor to perform functions including: (i) receiving, via the processor, the one or more properties of the fluid in the fluidic pathway detected by the sensor, and (ii) determining, based on the detected one or more properties of the fluid in the fluidic pathway, a presence of a separation gas in the fluid in the fluidic pathway.

Abstract: Flow cytometry investigation and flow cytometry result analysis to determine an optimized dilution factor range for flow cytometry investigation of a target sample fluid stock using a flow cytometer. The optimized dilution factor range may be determined using a screening assay module of a flow cytometry system to analyze flow cytometry results determined to fall within a dynamic range of a flow cytometer to determine the optimized dilution factor range for a given flow cytometer to investigate a given target sample fluid stock. In turn, a titer assay module may be executed to prepare particle titer results based on optimized target fluid samples provided within the optimized dilution factor range. The screening assay module and/or the titer assay module may provide automated data processing with data notifications and confirmations to provide robust data analysis.

Abstract: The disclosure provides example methods that include a processor: (a) generating at least one phase contrast image of a biological specimen comprising one or more cells centered around a focal plane for the biological specimen; (b) generating a confluence mask in the form of a binary image based on the at least one phase contrast image; (c) receiving a first brightfield image of the biological specimen at a defocusing distance above the focal plane and a second brightfield image of the biological specimen at the defocusing distance below the focal plane; (d) generating a cell image of the biological specimen based on the first and second brightfield image; (e) generating a seed mask based on the cell image and the phase contrast image; and (f) generating an image of the biological specimen showing a cell-by-cell segmentation mask based on the seed mask and the confluence mask.

Abstract: A method for evaluating a biological material for unassociated virus-size particles having an adenovirus 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-size particle and fluorescent antibody stain.

Abstract: A flow cytometer apparatus and methods for detecting and clearing a clog therein are disclosed. An example method for detecting a clog may include (i) detecting, via a fault detection system of a flow cytometer, a first plurality of events associated with a first aliquot from a first sample well, (ii) determining a count of the first plurality of events associated with the first aliquot, (iii) determining whether the count of the first plurality of events is below a minimum count tolerance and (iv) (a) if the count of the first plurality of events is below the minimum count tolerance, then determining that the flow cytometer has a clog, (b) if the count of the first plurality of events is equal to or above the minimum count tolerance, then detecting a second plurality of events associated with a second aliquot from a second sample well.

Abstract: Systems and methods are provided for microscopically and fluorescently imaging cell-bearing biological samples or other samples of interest. A microscope objective or other optical elements that exhibits chromatic aberration can be used to obtain images of fluorophores or other contrast agents at different wavelengths. The obtained images are then used to correct each other, e.g., to remove artifacts in an image of a shorter-wavelength fluorophore that are caused by cross-talk from a longer-wavelength fluorophore. A longer-wavelength image, taken at a focal distance corresponding to the shorter-wavelength fluorophore, is taken and used to subtract the activity of the longer-wavelength fluorophore in the shorter-wavelength image. The longer-wavelength image may be taken using a microscope set to the shorter-wavelength focal distance.

Abstract: A flow cytometry system and method for quantification of fluorescently labeled virus particles smaller than 1 micron in size measures and controls sample fluid flow rate to an investigatory flow cell at a rate below 5000 nanoliters per minute and calculates in real time particle concentration in the sample fluid using measured flow rated data flow rate data and with correction for sample fluid flow rate fluctuations.

Abstract: Systems, methods, and apparatuses of controlling fluid flow are disclosed. An apparatus includes a first microplate having a first open portion and defining one or more first wells therein, a second microplate having a second open portion and defining one or more second wells therein, and a pneumatic lid constructed of styrene ethylene butylene styrene (SEBS). The pneumatic lid extends over the first open portion and the second open portion and includes one or more microfluidic channels that fluidly couple the one or more first wells to the one or more second wells. The pneumatic lid provides an airtight seal over the first microplate and the second microplate.

Abstract: 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.

Abstract: A method for evaluating a biological material for unassociated virus-size particles of exosomes having a particular epitope uses a fluorescent antibody stain specific for binding with the epitope and a fluid sample with the exosomes 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 an exosome and fluorescent antibody stain.

Abstract: A method and a flow cytometer system in which a light path is aligned to optimally direct light from a light source to a flow cell of a flow cytometer instrument are provided. The flow cytometer may include an orientable mirror disposed in the light path between the light source and the flow cell. By changing an orientation of the mirror through a sequence of different orientations, an optimal orientation of the mirror may be determined, and the mirror may be oriented accordingly before proceeding to conduct a flow cytometry investigation.

Abstract: Methods of evaluating particle attributes in a sample fluid subjected to flow cytometry investigation in a flow cytometer instrument, methods of processing time series signal data traces output by a flow cytometer instrument, and a flow cytometer system are provided. In the methods and systems, data points comprising time series signal data traces corresponding with detection during the flow cytometry investigation of light from the sample fluid in one or more wavelength ranges indicative of the presence of one or more particle attributes in the sample fluid are batch-processed using a batch-specific signal peak threshold determined as a function of a batch-specific noise characteristic to identify signal peaks in the batch of data points indicative of the presence of the one or more particle attributes in the sample fluid.

Abstract: A method for detecting a separation gas in a fluid flow stream is provided herein. In one example, a voltage output signal is generated by a scatter detector of a flow cytometer as a flow stream of a plurality of gas-separated samples passes through the flow cytometer. The voltage output signal is sampled, and a timestamp and a voltage value are recorded for each sampled voltage of the voltage output signal that is greater than a separation gap threshold. In some examples the separation gap threshold is at least two times greater than a maximum voltage output of the samples. Flow cytometry systems including software configured to perform these method steps are also described.

Abstract: A method for evaluating a biological material for unassociated virus-size particles 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-size particle and fluorescent antibody stain.

Abstract: Apparatus and methods to improve the Boyden chamber used in cellular biological measurements, allowing quantitative optical microscopy of biological cells in situ without using fluorescent probes or optical staining. In the preferred embodiment, a thin porous membrane separating top and bottom reservoirs includes an array of precisely positioned micropores pores manufactured using a laser-based photo-machining (ablation) process. The membrane may be composed of polyethylene terephthalate (PET), polycarbonate, polyimide, polyether ether ketone (PEEK) or other appropriate material. The pores formed in the membrane may have diameters in the range of 1 to 15 microns and spaced apart at a distance ranging from 10 to 200 microns. A plurality of upper and lower reservoirs may be provided to form a multi-well plate.

Abstract: Apparatus and methods to improve the Boyden chamber used in cellular biological measurements, allowing quantitative optical microscopy of biological cells in situ without using fluorescent probes or optical staining. In the preferred embodiment, a thin porous membrane separating top and bottom reservoirs includes an array of precisely positioned micropores pores manufactured using a laser-based photo-machining (ablation) process. The membrane may be composed of polyethylene terephthalate (PET), polycarbonate, polyimide, polyether ether ketone (PEEK) or other appropriate material. The pores formed in the membrane may have diameters in the range of 1 to 15 microns and spaced apart at a distance ranging from 10 to 200 microns. A plurality of upper and lower reservoirs may be provided to form a multi-well plate.

Abstract: Systems, including apparatus and methods, for performing electrophysiological measurements on membranous samples, including living cells, isolated cell fragments (such as organelles), and/or artificial membranes (such as vesicles). The apparatus may include a high-throughput electrophysiological measurement system, and components thereof. This measurement system may include, among others, (1) a fluidics head for transferring samples and/or other compounds to a perforated measurement substrate, (2) a pressure-regulated plenum system for positioning samples on the substrate and subsequently forming a high-resistance electrical seal, (3) an activation system (such as a computer-controlled pulsed UV illumination module) for activating caged compounds, (4) an electronics head for applying and/or measuring voltage and/or current, and/or (5) a computer-controlled analysis system for collecting and/or analyzing data.