Indevr, Inc.

Intellicyt Corporation

Essen Instruments, Inc.

University of Colorado

University Technology Corporation

Virocyt, Inc.

Kathy Lou Rowlen

Erica Dawson Tenent

Andrew Smolak

Jean-Luc Fraikin

Christopher Bowman

Christopher Converse

John William Birks

Garrett S Wilson

Hadley Sikes

Ryan Hansen

Matthew Ferris

Eric D Smith

Robert Stoughton

Craig Joseph Williford

Evan M Toth

Lauren R Wolfe

Goran Rauker

Heather Jean Avens

Jeffrey T Ives

Robert Kuchta

Laura R Kuck

Sarah G Westerbuhr

Garrett Wilson

Gregory Yamada

Provided herein are enhanced imaging techniques which allow for the use of multiple optical sensors, each of which corresponds to only a portion of an array, including utilizing individual sensors for each individual well of a multi-well plate. The provided systems and methods may reduce the amount of time to perform optical analysis of an array, reduce the amount of moving parts or mechanical devices required to perform optical analysis and/or reduce the amount of space between the sensors and the array being analyzed resulting in more compact, efficient optical analyzers.

Parallel imaging system

The systems and methods contained herein are directed toward automated analysis of agglutination reactions to determine properties of materials, including viruses and vaccines thereto. Advanced digital imaging and processing techniques are used to determine the presence or absence of viruses or antibodies within a fluid sample. The systems and methods are versatile, and can be used to determine specific properties of biomaterials and viruses, such as titer value, concentration, genotype, phenotype, serotype, vaccine efficacy, viral resistance and other properties of relevance in the medical, research and development fields. Also provided are systems and methods of standardization, repeatability, and data storage and transmittal to reduce errors and subjectivity inherent to conventional assays characterized by human readers.

Automated agglutination analyzer with contour comparison

A universal array for multiplexed quantification of variable hemagglutinin such as subcomponents of multivalent annual influenza vaccines that is robust to variations in proteins such as mutations and is capable of quantifying degradation of proteins. Universal capture array () comprises one or more substrates () and a low-density microarray () of sub-arrays () comprising spots (-). The microarray () is contacted with one or more targets () at one or more unknown concentrations, and bound complexes () are formed and subsequently quantified with a suitable method. Quantified signals are compared to calibration curves to obtain one or more unknown concentrations and/or quantify degradation of the one or more targets (). Other embodiments are described and shown.

Universal capture array for multiplexed subtype-specific quantification and stability determination of influenza proteins

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.

Flow measurement and control for improved quantification of particles in flow cytometry

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.

Automated alignment of optics within a flow cytometer

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.

Flow cytometer signal peak identification employing dynamic thresholding

Methods for the quantification of influenza HA proteins and anti-influenza antibodies for the fields of vaccine-related protein quantification, potency determination, and efficacy evaluation are provided. According to the technology, quantification is achieved by providing capture agents attached to an array in a series of decreasing concentrations. Serial dilutions of a reference material also may be introduced. The reference material within each solution binds to the capture agents on the array and is labeled with a label agent capable of producing a detectable signal used to construct a calibration curve. A target material of unknown concentration is introduced to a separate identical array, and the target material binds to the capture agents and also is labeled by a label agent to produce a detectable signal. The calibration curve based on the reference material is then utilized to determine the concentration of the target material without the need to perform replicate experiments.

Low density microarrays for vaccine related protein quantification, potency determination and efficacy evaluation

The present invention relates to methods that enable improved accuracy for quantitative particle counting in a flowing liquid stream. The methods of the present invention utilize the real-time measurement of flow rates and flow rate control through feedback mechanisms to improve quantification, and this improved quantification translates to more accurate particle counting. In certain embodiments, particles being counted are biological particles in a liquid sample, such as viruses.

A flow cytometer may include a vibration isolation structure on which a flow cytometer optical system assembly is supported when the flow cytometer is in an operational configuration. A shear protection structure may be positioned to protect a vibration isolation structure from damage during handling and shipping. A flow cytometer optical system assembly may include optical component units fixed in position on a support platform with adjustability of one or more optical features in the optical component units. A light-tight dichroic mirror unit may include a rotatably mounted dichroic mirror with a locking mechanism to permit re-setting angular positioning of a dichroic mirror.

Flow cytometer with optical system assembly having vibration isolation and shear protection structures

In a method for processing biological materials for flow cytometry evaluation for virus particles, a mixture including biological material and purification particles is centrifuged to prepare a centrifuged composition including a supernatant that may be further processed prior to the flow cytometry evaluation. The purification particles include porous cores functionalized to capture smaller-size impurities in a biological material sample and a porous size-exclusion shell surrounding the core to exclude larger-size components of the biological material from entering into the core. Multiple samples may be processed in multi-sample processing units. A product may contain a sealed container with the unit quantity of purification particle in a storage liquid and a kit may include such a sealed container and a centrifugal filter.

Processing biological material for flow cytometry evaluation for virus particles

A system for the indirect detection of agglutination in an agglutination assay using optical density measurement and a reaction vessel. Light emitted by light source is transmitted through reaction vessel containing components of an agglutination reaction that is detected by light detector. As particles in the reaction vessel settle in the absence of agglutination they are concentrated in a reaction vessel lower volume and increase the optical density of the light path through the reaction vessel. Light detector generates an output signal related to said optical density. Said output signal is read and interpreted by an instrument to detect a presence or absence of agglutination. Other embodiments are described and shown.

Detection of agglutination by optical density measurement

The invention provides methods to detect molecular recognition events. The invention also provides methods to detect the presence of or identify a target species based on its interaction with one or more probe species. The methods of the invention are based on amplification of the signal due to each molecular recognition event. The amplification is achieved through photopolymerization, with the polymer formed being associated with the molecular recognition event. In one aspect, a fluorescent polymer, a magnetic polymer, a radioactive polymer or an electrically conducting polymer can form the basis of detection and amplification. In another aspect, a polymer gel swollen with a fluorescent solution, a magnetic solution, a radioactive solution or an electrically conducting solution can form the basis of detection and amplification. In another aspect, detectable particles can be included in the polymer formed. In another aspect, sufficient polymer forms to be detectable by visual inspection.

Use of photopolymerization for amplification and detection of a Molecular Recognition Event

The invention provides methods to detect molecular recognition events. The invention also provides methods to detect the presence of or identify a target species based on its interaction with one or more probe species. The methods of the invention are based on amplification of the signal due to each molecular recognition event. The amplification is achieved through photopolymerization, with the polymer formed being associated with the molecular recognition event. In an embodiment, a fluorescent polymer, a magnetic polymer, a radioactive polymer or an electrically conducting polymer can form the basis of detection and amplification. In another embodiment, a polymer gel swollen with a fluorescent solution, a magnetic solution, a radioactive solution or an electrically conducting solution can form the basis of detection and amplification. In another embodiment, sufficient polymer forms to be detectable by visual inspection.

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Longmont, CO, United States of America

Kathy Lou Rowlen