Qvella Corporation

Novx Systems Inc.

Other

Novx Systems Canada, Inc.

Stephen Wesley Leonard

Samad Talebpour

Tino Alavie

Robert Maaskant

Aye Aye Khine

Cyrus Robert Etemad-Moghadam

Ming Gang Xu

Sanjesh Yasotharan

Alexander Zahn

Keith Beckley

Ulrich Gosele

Ralf Wehrspohn

Henry M Van Driel

Stefan Senz

Maryam Asadishekari

Anna Khimchenko

Andrew Hudson

William Yang

Jorg Schilling

Systems and methods are provided for classifying microbial cells according to morphological features of microcolonies. A dark-field objective is employed to acquire a dark-field image of a microcolony during a microcolony growth phase that is characterized by phenotypic expression of microcolony morphological features which evolve with time and are differentiated among classes of microbial cell types. The dark-field image is processed to classify the microcolony according to two or more microbial cell types, such as Gram status and/or speciation. The dark-field objective may have a numerical aperture selected to facilitate the imaging of microcolony morphological features, residing, for example, between 0.15 and 0.35. A set of dark-field images of a microcolony may be collected during the microcolony growth phase and processed to classify the microcolony. Classification may be performed according to a temporal ordering of the dark-field images, for example, using a recurrent neural network.

Systems and methods for classification of microbial cells grown in microcolonies

Methods are provided for performing antibiotic susceptibility testing based on the detection of RNA, such as tmRNA, from microbial cells after exposure to antibiotics. In some embodiments, aliquots are obtained from a sample, one of which contains a selected antibiotic. The aliquots, which include growth media, are incubated under conditions suitable for microbial growth, and the microbial cells in each aliquot are removed and lysed, and the lysate is subjected to reverse transcription and amplification in infer the effect of the selected antibiotic on the microbial cells. In one embodiment, a sample containing microbial cells is incubated in the presence of a selected antibiotic and a stimulus is provided to induce the production of m RNA within the microbial cells. The microbial cells are subsequently lysed without substantial degradation of the m RNA within the lysate, and the m RNA is detected to determine the effect of the antibiotic on the microbial cells.

Methods of targeted antibiotic susceptibility testing

Systems, methods and devices are provided for the automated centrifugal processing of samples. In some embodiments, an integrated fluidic processing cartridge is provided, in which a centrifugation chamber is fluidically interfaced, through a lateral surface thereof, with a microfluidic device, and wherein the integrated fluidic processing cartridge is configured to be inserted into a centrifuge for centrifugation. A cartridge interfacing assembly may be employed to interface with the integrated fluidic processing cartridge for performing various fluidic processing steps, such as controlling the flow of fluids into and out of the centrifugation chamber, and controlling the flow of fluids into the microfluidic device, and optionally for the further fluidic processing of fluids extracted to the microfluidic device. The integrated fluidic processing cartridge may include a supernatant chamber the extraction of a supernatant thereto, and a diluent chamber for diluting a suspension collected in the centrifugation chamber.

Apparatus, system and method for performing automated centrifugal separation

The present invention provides a microfluidic devices and methods of use thereof for the concentration and capture of cells. A pulsed non Faradaic electric field is applied relative to a sample under laminar flow, which results to the concentration and capture of charged analyte. Advantageously, pulse timing is selected to avoid problems associated with ionic screening within the channel. At least one of the electrodes within the channel is coated with an insulating layer to prevent a Faradaic current from flowing in the channel. Under pulsed application of a unipolar voltage to the electrodes, charged analyte within the sample is moved towards one of the electrodes via a transient electrophoretic force.

Cell concentration, capture and lysis devices and methods of use thereof

Systems, methods and devices are provided for the automated centrifugal processing of samples. In some embodiments, an integrated fluidic processing cartridge is provided, in which a centrifugation chamber is fluidically interfaced, through a lateral surface thereof, with a microfluidic device, and wherein the integrated fluidic processing cartridge is configured to be inserted into a centrifuge for centrifugation. A cartridge interfacing assembly may be employed to interface with the integrated fluidic processing cartridge for performing various fluidic processing steps, such as controlling the flow of fluids into and out of the centrifugation chamber, and controlling the flow of fluids into the microfluidic device, and optionally for the further fluidic processing of fluids extracted to the micro-fluidic device. The integrated fluidic processing cartridge may include a supernatant chamber the extraction of a supernatant thereto, and a dilutent chamber for diluting a suspension collected in the centrifugation chamber.

Methods are provided for performing antibiotic susceptibility testing based on the detection of RNA, such as tmRNA, from microbial cells after exposure to antibiotics. In some embodiments, aliquots are obtained from a sample, one of which contains a selected antibiotic. The aliquots, which include growth media, are incubated under conditions suitable for microbial growth, and the microbial cells in each aliquot are removed and lysed, and the lysate is subjected to reverse transcription and amplification in infer the effect of the selected antibiotic on the microbial cells. In one embodiment, a sample containing microbial cells is incubated in the presence of a selected antibiotic and a stimulus is provided to induce the production on m RNA within the microbial cells. The microbial cells are subsequently lysed without substantial degradation of the m RNA within the lysate, and the m RNA is detected to determine the effect of the antibiotic on the microbial cells.

The present invention provides a microfluidic devices and methods of use thereof for the concentration and capture of cells. A pulsed non-Faradaic electric field is applied relative to a sample under laminar flow, which results to the concentration and capture of charged analyte. Advantageously, pulse timing is selected to avoid problems associated with ionic screening within the channel. At least one of the electrodes within the channel is coated with an insulating layer to prevent a Faradaic current from flowing in the channel. Under pulsed application of a unipolar voltage to the electrodes, charged analyte within the sample is moved towards one of the electrodes via a transient electrophoretic force.

Assays and methods of compensating for changes in a dose-response curve of an assay due to variations in a perturbing variable by firstly measuring the dose-response curve, the individual assay parameters, at different values of the perturbing variable. Next, unknown samples are assayed simultaneously with a known standard at a chosen analyte concentration with a value of the perturbing variable being unknown and the dose-response curve unknown. The different dose-response curves from the first step are used to determine a mathematical relationship between assay parameters and the assay signal of the known standard. The assay parameters that are valid for the unknown value of the perturbing variable can be obtained by substituting the value of the assay signal from the known standard into the mathematical relationship and solving for the assay parameters.

Method of compensation of dose-response curve of an assay for sensitivity to perturbing variables

The present invention provides assays and methods of compensating for changes in an assay where such changes are due to variations in a perturbing variable. This is achieved by a two-step method, the first step of which involves measurements of the dose-response curve, and thus the individual assay parameters, at different values of the perturbing variable. In the second step, unknown samples are assayed simultaneously with a standard. During this measurement, the value of the perturbing variable is unknown and the dose-response curve is therefore also unknown. The dose-response curves from the first step are used to determine a mathematical relationship between the assay parameters and the assay signal of the standard. Assay parameters that are valid for the unknown value of the perturbing variable can be obtained by substituting the value of the assay signal from the standard into the mathematical relationship and solving for the assay parameters.

The present invention provides a method of performing a competitive assay for the detection and quantification of an analyte over an extended dynamic range. This is achieved by a multi-step sample addition method whereby different concentrations of sample are added at different times during the assay that produces a dose-response curve with multiple windows of detection. This multi-step sample addition method causes the dose-response curve of the composite assay to broaden, dramatically increasing the assay dynamic range.

Immunoassay with extended dynamic range

The present invention provides a bar-code driven, completely automated, microplate-based analyzer system for performing chemical, biochemical or biological assays. The analyzer is a modular, bench-top instrument that compactly integrates subsystems for sample dispensing, liquid handling, microplate transport, thermal incubation, vortexing, solid phase separation and optical reading. An internal processor is included for automating the instrument, and a user interface to facilitate communication with the operator via a touch-sensitive liquid-crystal display (LCD), and communicating with a remote network via multiple protocols. The analyzer includes firmware resident within the processing system and the user interface allows the operator to select pre-defined assay batch protocols and the user interface is configured in such as way so as to restrict an operator from programming the firmware.

Automated analyzer

An optical system is provided for achieving enhanced rejection of scattered excitation light and superior signal-to-noise performance when reading microplate wells. The optical system uses an axial configuration in which the excitation beam incident upon the sample propagates along the axis of the microplate well. Excitation light from a light source, such as a lamp or fiber optic bundle, is collimated into a beam using a lens. A reflective pick-off mirror is then used to reflect the collimated excitation beam upward along the well axis. A focusing lens, with a diameter exceeding the diameter of the collimated excitation beam, is used to focus the excitation beam in the well. The same broad lens is used to collimate the emitted fluorescent light, of which a large percentage propagates axially past the pick-off mirror towards a second focusing lens that focuses the emission beam onto the face of a fiber optic bundle. The emitted light is later filtered and detected using at a position that is optically shielded from the aforementioned optical system. The optical system is incorporated into a microplate reader or automated assay instrument in order to provide a compact assembly for sensitive fluorescence measurements either above or below the microplate. The optical system further enables the simultaneous measurement of absorbance and fluorescence in a compact optical configuration.

Growing community of inventors

Unionville, Canada

Stephen Wesley Leonard