US Pat. No. 9,376,718

METHOD AND APPARATUS FOR GENERATING THERMAL MELTING CURVES IN A MICROFLUIDIC DEVICE

Caliper Life Sciences, In...

1. A method of performing thermal melt analysis of a nucleic acid in a microfluidic device, the method comprising:
providing a microfluidic device having at least one microfluidic channel,
introducing fluid comprising the nucleic acid and amplification reagents into the microfluidic channel so that the fluid continuously
flows through the channel,

cycling the temperature in at least one portion of the microfluidic channel so that the nucleic acid undergoes amplification,
wherein the cycling of the temperature comprises varying the temperature of the at least one portion using a non-joule heating
method,

after the amplification is completed, subjecting the fluid to a continuously increasing series of temperatures in the same
microfluidic channel, wherein the series of temperatures includes a temperature high enough to cause denaturation of the nucleic
acid, and

measuring a detectable property emanating from the fluid in the same microfluidic channel while subjecting the fluid to the
continuously increasing series of temperatures, wherein the detectable property is indicative of the extent of denaturation
of the nucleic acid.

US Pat. No. 9,283,563

SYSTEMS AND METHODS FOR REAL-TIME PCR

Canon U.S. Life Sciences,...

1. A real-time DNA amplification and analysis method, comprising:
creating a script, wherein the script contains configuration information;
preparing a microfluidic device having a microfluidic channel having a PCR zone and a DNAmelting zone;
reading the script to obtain the configuration information;
positioning a well plate relative to the device, the well plate having a buffer well containing a buffer solution and sample
well containing a sample solution containing a DNA sample;

activating a pump, wherein the pump is configured to create a pressure differential that causes the sample solution to flow
through the channel, wherein the sample flows through the PCR zone prior to flowing through the DNA melting zone;

while the sample is flowing through the channel:
cycling the temperature of the sample according to configuration information included in the script as the sample flows through
the PCR zone to amplify the DNA sample;

illuminating the sample with a first excitation source as the sample flows through the PCR zone and using a first image detector
to obtain more than one image of the sample;

processing the images;
determining whether another PCR cycle is needed, and if so, causing additional temperature cycling to occur; and
melting the amplified DNA, illuminating the sample with a second excitation source as the sample flows through the DNA melting
zone and using a second image detector to obtain more than one image of the sample, and wherein the first and second excitation
sources are of different types.

US Pat. No. 9,114,399

SYSTEM AND METHOD FOR SERIAL PROCESSING OF MULTIPLE NUCLEIC ACID ASSAYS

Canon U.S. Life Sciences,...

1. An instrument for serial processing of multiple nucleic acid assays, comprising:
a frame chassis;
a processing drawer configured to be moveable relative to the frame chassis between an open position and a closed position
and including a microfluidic device support structure configured to hold a microfluidic device;

a cooling manifold assembly carried on a portion of said frame chassis adjacent said processing drawer and configured to be
in an operative position with respect to the microfluidic device support structure when the processing drawer is in the closed
position, wherein the cooling manifold assembly is connected to an upper surface of the microfluidic device, the cooling manifold
assembly is configured to direct airflow to the microfluidic device supported on the microfluidic device support structure
while isolating the airflow from one or more inlet ports of the microfluidic device, and the cooling manifold assembly comprises
one or more openings extending through the cooling manifold assembly to access the one or more inlet ports of the microfluidic
device, and the rest of the microfluidic device is covered by the cooling manifold;

a liquid handling system supported by the frame chassis, wherein the liquid handling system is configured such that the microfluidic
device mounted on the microfluidic device support structure is accessible to the liquid handling system when the processing
drawer is in the closed position; and

an optical imaging system configured to create images of fluorescent emissions from materials within a microfluidic channel
of the microfluidic device, the optical imaging system being carried on a portion of said frame chassis adjacent said processing
drawer and configured to be in an operative position with respect to the microfluidic device support structure when the processing
drawer is in the closed position.

US Pat. No. 9,492,826

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. In a microfluidic device incorporating thin-film heaters within a plurality of microfluidic channels to effect a biological
reaction with multiple steps each requiring a different temperature, a method of controlling the microfluidic with pulse width
modulation (PWM) comprising:
generating a set of calibration data while calibrating a modulation frequency with a temperature by measuring a cooling rate
of the thin-film heaters between pulses and adjusting the modulation frequency based upon the measured rate;

using the calibration data to generate a PWM control signal for the thin-film heaters designed to bring the heaters to a desired
temperature for a first step in the biological process;

adjusting the PWM signal to achieve a temperature for an appropriate time required by said first step; and
adjusting the PWM signal to achieve a desired temperature for a second step in the biological process.
US Pat. No. 9,116,088

METHODS AND SYSTEMS FOR DNA ISOLATION ON A MICROFLUIDIC DEVICE

Canon U.S. Life Sciences,...

1. A method of isolating DNA from cells in a sample comprising the steps of:
(a) selectively lysing the cellular membranes of the cells in the sample without lysing the nuclear membranes of the cells
to produce intact nuclei from the cells;

(b) separating the intact nuclei from the sample by a nuclei size exclusion barrier in a nuclei separation region of a microfluidic
device by flowing the sample through the nuclei size exclusion barrier in a first direction;

(c) flowing an elution buffer in a second direction into the nuclei separation region of the microfluidic device to resuspend
the isolated nuclei; wherein the first direction is substantially orthogonal to the second direction;

(d) delivering the resuspended isolated nuclei to a nuclei lysis region of the microfluidic device; and
(e) lysing the resuspended isolated nuclei in the nuclei lysis region of the microfluidic device to release the isolated DNA.

US Pat. No. 9,554,422

SYSTEMS AND METHODS USING EXTERNAL HEATER SYSTEMS IN MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. A heating system for microfluidic devices comprising:
a) a microfluidic device having two or more reservoirs or channels;
b) a heat spreader, wherein the heat spreader is affixed to the microfluidic device such that the reservoirs or channels disposed
on said microfluidic device are in thermal communication with the heat spreader, wherein the heat spreader is made of anisotropic
material and is aligned with the microfluidic device to provide uniformity of temperature between the two or more channels,
wherein a high conductance orientation of the heat spreader is aligned parallel to the plane having the two or more reservoirs
or channels;

c) a heating means for heating the heat spreader;
d) a measuring means for measuring one or more temperatures of the channels or reservoirs, wherein the measuring means comprises
one or more temperature sensors; and,

wherein (i) an external resistive heater and an external temperature sensor are attached to the heat spreader and (ii) the
microfluidic device comprises at least one embedded temperature sensor.

US Pat. No. 9,427,736

SYSTEM AND METHOD FOR MICROFLUIDIC FLOW CONTROL

Canon U.S. Life Sciences,...

1. A system for sequentially introducing predetermined amounts of different reaction fluids into a microfluidic circuit comprising:
a. a microfluidic circuit comprising a microchannel, a plurality of inlet ports in fluid communication with the microchannel
and through which different reaction fluids are introduced into the microchannel and an outlet port in fluid communication
with the microchannel through which fluid from the microchannel is collected;

b. at least one pressure source in selective communication with said outlet port and said plurality of inlet ports; and
c. at least one valve mechanism operatively associated with said plurality of inlet ports and in communication with said pressure
source, each valve mechanism being adapted to sequentially connect each of said plurality of inlet ports to a pressure generated
by said pressure source for a predetermined period of time, causing a predetermined amount of fluid to flow from the inlet
port into the microchannel, and then connecting said inlet port to a different pressure to substantially stop the flow of
fluid.

US Pat. No. 9,404,152

MICROFLUIDIC FLOW MONITORING

Canon U.S. Life Sciences,...

1. A method of monitoring a solution flow in a microfluidic channel comprising:
(a) moving a solution through the microfluidic channel in a chip to fill the channel with a solution and stop the solution
flow to thermally cycle the entire length of the channels to perform a polymerase chain reaction (PCR) on the solution flow
when the solution flow is stopped;

(b) introducing a flow marker into the channel;
(c) illuminating the channel at two measuring points; and
(d) measuring the movement of the flow marker within the channel between the two measuring points.

US Pat. No. 9,061,278

MICROFLUIDIC SYSTEMS AND METHODS FOR THERMAL CONTROL

Canon U.S. Life Sciences,...

1. A method for individually controlling a plurality of resistive thermal detectors (RTDs) of a microfluidic device of a microfluidic
system, wherein the RTDs are each adjacent to a portion of an associated one of the plurality of microchannels, the method
comprising:
generating heater control signals having alternating polarities to drive the plurality of RTDs;
supplying the heater control signals to the plurality of RTDs so that adjacent RTDs of the plurality of RTDs are driven with
heater control signals having opposite polarities relative to each other;

minimizing current in first and second common electrodes by connecting the first and second common electrodes to a virtual
ground circuit, wherein the first and second common electrodes are each connected to each RTD of the plurality of RTDs;

sensing a temperature of each of the plurality of RTDs by using the first and second common electrodes; and
updating the heater control signals using the sensed temperatures of the plurality of RTDs.

US Pat. No. 9,447,458

DETECTION OF NEIGHBORING VARIANTS

Canon U.S. Life Sciences,...

1. A method of distinguishing between at least two nearby neighbor variants on a locus of interest on a gene, the method comprising:
(a) providing a first aliquot of said nucleic acid having the locus of interest;
(b) incubating said first aliquot of said nucleic acid with a limiting primer, an excess primer, and a first probe that is
designed to hybridize to said locus of interest on a target strand of said nucleic acid;

(c) performing asymmetric PCR using said first aliquot to produce an excess of amplicons corresponding to the target strand
to which the first probe hybridizes, thereby producing a first probe element;

(d) providing a second aliquot of said nucleic acid target having the locus of interest;
(e) incubating said second aliquot of said nucleic acid target with said limiting primer, said excess primer, and a second
probe that is designed to hybridize to said locus of interest on the target strand, wherein said first probe differs in sequence
from said second probe in length;

(f) performing asymmetric PCR using said second aliquot to produce an excess of amplicons corresponding to the target strand
to which the second probe hybridizes, thereby producing a second probe element, wherein said first probe and said second probe
each have a sequence that is complementary to a wild-type sequence of the target strand, each of said first and second probes
covering the same at least two nearby neighbor variants;

(g) generating a first melting curve for the first probe element in a first mixture with a saturating binding dye by measuring
fluorescence from said dye as the first mixture is heated;

(h) generating a second melting curve for the second probe element in a second mixture with said saturating binding dye by
measuring fluorescence from said dye as the second mixture is heated; and

(i) analyzing said first melting curve and said second melting curve to distinguish between said at least two nearby neighbor
variants, wherein a melting signature curve of each of said at least two nearby neighbor variants is different in said first
and second melting curves.

US Pat. No. 9,267,852

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A method for determining the temperature of each of a plurality of multiplexed heater electrodes, wherein the heater electrodes
are part of a multiplex circuit sharing a common lead connecting the electrodes to a power supply, which multiplex circuit
includes a controller, said method comprising:
a. with the power supply connected to the common lead, independently measuring a voltage drop of each heater electrode in
series with the common lead and storing, by the controller, a common power voltage drop data for each of the heater electrodes;

b. disconnecting, by the controller, the power supply from the common lead;
c. connecting, by the controller, the power supply to each of one or more of the heater electrodes, wherein the power supply
is connected to one or more of the heater electrodes at a time;

d. while the power supply is connected to a heater electrode, isolating, by the controller, at least one other heater electrode
from all other heater electrodes of the multiplex circuit except the heater electrode connected to power supply, measuring
an isolated voltage drop at each isolated heater electrode, and storing, by the controller, isolated voltage drop data for
each isolated heater electrode;

e. computing, by the controller, the resistance of each of the plurality of multiplexed heater electrodes by solving for the
resistance of each heater electrode based at least in part on the stored common power voltage drop data and the stored isolated
voltage drop data; and

f. deriving, by the controller, the temperature of each of the plurality of multiplexed heater electrodes from the computed
resistance of each electrode.

US Pat. No. 9,278,321

CHIP AND CARTRIDGE DESIGN CONFIGURATION FOR PERFORMING MICRO-FLUIDIC ASSAYS

Canon U.S. Life Sciences,...

1. A system for performing microfluidic assays, the system comprising:
a microfluidic chip comprising a DNA amplification area and an analysis area in communication with at least a first access
port and a second access port; and

a cartridge device configured to removably interface with the micro-fluidic chip, the cartridge device comprising:
a delivery chamber in fluid communication with a delivery port, wherein said delivery chamber is configured to contain a reaction
fluid and said delivery port is configured to removably interface with the first access port of the micro-fluidic chip; and

a recovery chamber in fluid communication with a recovery port, wherein said recovery chamber is configured to receive waste
materials from the second access port of said micro-fluidic chip and said recovery port is configured to removably interface
with said micro-fluidic chip;

wherein the cartridge device delivers fluids to and removes fluids from the microfluidic chip, wherein a connection between
the microfluidic chip and the cartridge device is limited to coupling the delivery port to the first access port and the recovery
port to the second access port; and,

wherein the DNA amplification area and the analysis area of the microfluidic chip are configured to support an amplification
reaction and a subsequent analysis.

US Pat. No. 9,221,056

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A method for controlling the temperature of a heater electrode associated with a microfluidic channel of a microfluidic
device, wherein power applied to the heater electrode is regulated by varying the duty cycle of a pulse width modulation (PWM),
and wherein the method comprises:
a. applying a fixed voltage across a circuit including the heater electrode, a switching element for selectively opening or
closing the circuit to define the duty cycle, and a high resistance shunt around the switching element;

b. closing the circuit with the switching element for a period time corresponding to a power-on portion of a desired duty
cycle and passing current to the heater electrode through the closed switch;

c. while the circuit is closed, measuring a power-on voltage drop across the heater electrode;
d. computing a power-on resistance of the heater electrode based on the fixed voltage, the measured power-on voltage drop,
and the known resistance of the circuit including the resistance of the shunt and the resistance of a branch of the circuit
including the switch;

e. deriving a power-on temperature of the heater electrode from the computed power-on resistance;
f. opening the circuit with the switching element for period of time corresponding to a power-off portion of the desired duty
cycle, and passing current to the heater electrode exclusively through the shunt;

g. while the circuit is opened, measuring a power-off voltage drop across the heater electrode;
h. computing a power-off resistance of the heater electrode based on the fixed voltage, the measured power-off voltage drop,
and the known resistance of the circuit including the resistance of the shunt but not the resistance of the branch of the
circuit including the switch;

i. deriving a power-off temperature of the heater electrode from the computed power-off resistance; and
j. comparing at least one of the power-on temperature and the power-off temperature to a desired temperature of the heater
electrode and adjusting the duty cycle if necessary to reduce the difference between the power-on or power-off temperature
and the desired temperature.

US Pat. No. 9,272,282

COMBINED THERMAL DEVICES FOR THERMAL CYCLING

Canon U.S. Life Sciences,...

1. A method for heating a nucleic acid sample in a microfluidic device by using a heating device and an electromagnetic heating
device, the method comprising:
(a) controlling the heating device to cause a temperature of the sample to be at or about a first desired temperature for
at least a first time period;

(b) after expiration of the first time period, increasing the output of the electromagnetic heating source to cause the temperature
of the sample to be at or about a second desired temperature for at least a second time period;

(c) during said second time period, lowering the amount of heat the heating device provides to the sample; and
(d) immediately after expiration of the second time period, lowering the output of the electromagnetic heating source and
controlling the heating device to cause the temperature of the sample to be at or about a third desired temperature for a
third time period,

wherein the first temperature is less than the second temperature and the third temperature is less than the first temperature,
wherein the temperature of the sample during the second time period is caused by heat provided by both the heating device
and the electromagnetic heating source heating the nucleic acid sample.

US Pat. No. 9,393,566

SYSTEM AND METHOD FOR TEMPERATURE REFERENCING FOR MELT CURVE DATA COLLECTION

Canon U.S. Life Sciences,...

1. A method of performing melt curve data collection comprising:
(a) providing a container which comprises at least two chambers that are in close thermal connection;
(b) introducing a DNA sample to be tested into at least one of the chambers and a temperature reference material exhibiting
a known optical property change as a function of temperature into at least one of the other chambers, wherein the temperature
reference materials bracket the DNA sample in space;

(c) heating the chambers from a first temperature to a second temperature;
(d) simultaneously measuring a detectable property emanating from the DNA sample and a detectable property emanating from
the temperature reference material during step (c), wherein the detectable property of the DNA sample indicates an extent
of denaturation of the DNA in the sample and the detectable property of the temperature reference material is correlated to
the temperature of the chamber where the temperature reference material is located; and,

(e) comparing the detectable property of the DNA sample with the detectable property of the temperature reference material
to determine the actual melt properties of the DNA sample, wherein the detectable property emanating from the temperature
reference materials is measured to determine a spatial temperature gradient and any temporal fluctuation and a temperature
at the location of the DNA sample is estimated by interpolating the results from temperature reference materials that surround
the location.

US Pat. No. 9,114,397

METHOD OF REDUCING CROSS-CONTAMINATION IN CONTINUOUS AMPLIFICATION REACTIONS IN A CHANNEL

Canon U.S. Life Sciences,...

1. A method of performing continuous flow amplification reactions in a microfluidic channel with reduced cross-contamination,
the method comprising:
(a) introducing a plug of a sample solution into a microfluidic channel, then introducing a plug of a cleaning solution into
the microfluidic channel, whereby the plugs are introduced in a continuous flow such that the plugs of sample solution alternate
with the plugs of cleaning solution in the microfluidic channel, wherein the sample solution comprises MgCl2 and dNTPs, and wherein the cleaning solution comprises one or more zwitterions, wherein the plug of the sample solution and
the plug of the cleaning solution are sequentially introduced into the microfluidic channel through the same inlet port;

(b) introducing a solution comprising a primer and a solution comprising a polymerase into each plug of the sample solution
as each plug flows through a portion of the micro fluidic channel; and

(c) performing amplification reactions on the plugs of sample solutions as they continuously flow through the microfluidic
channel,

wherein the cleaning solution reduces MgCl2 adherence to the microfluidic channel surface; and wherein cross-contamination of nucleic acids in the amplification reactions
is substantially eliminated.

US Pat. No. 9,292,653

HIGH-RESOLUTION MELTING ANALYSIS

Canon U.S. Life Sciences,...

1. A method for identifying one or more nucleic acids in a sample, the method comprising:
an imaging system obtaining thermal dissociation data for the one or more nucleic acids by measuring a signal emanating from
the sample as the temperature of the sample is increasing over a nucleic acids dissociation range, wherein the signal is indicative
of a nucleic acid denaturation;

a processor in communication with the imaging system providing a mathematical representation to model the thermal dissociation
data, the mathematical representation being a sum including one or more terms, wherein each of the one or more terms is associated
with one of the nucleic acids in the sample, wherein each of the one or more terms depends on the temperature of the sample
and parameters defining the one of the nucleic acids in the sample;

fitting by the processor the measured thermal dissociation data to the mathematical representation to calculate the parameters
defining each of the one or more nucleic acids in the sample; and

identifying by the processor each of the one or more nucleic acids in the sample based on the calculated parameters.

US Pat. No. 9,168,529

AIR COOLING SYSTEMS AND METHODS FOR MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. An instrument comprising:
a microfluidic device including:
one or more microfluidic channels, one or more inlet ports configured to introduce biological material into the one or more
microfluidic channels, one or more outlet ports, and one or more heat sinks positioned above the one or more microfluidic
channels, wherein the one or more heat sinks and the one or more inlet and outlet ports are located on a first side of the
microfluidic device;

a cooling manifold configured to direct an airflow to the one or more heat sinks of the microfluidic device while the cooling
manifold is attached to the first side of the microfluidic device, wherein said cooling manifold is configured to isolate
the airflow from the one or more inlet ports, the cooling manifold including:

an air inlet configured to receive the airflow;
an air outlet; and
an inlet duct configured to direct the airflow from the air inlet to the one or more heat sinks of the microfluidic device.
US Pat. No. 9,114,398

DEVICE AND METHOD FOR DIGITAL MULTIPLEX PCR ASSAYS

Canon U.S. Life Sciences,...

1. A method for performing digital multiplex PCR assay within a plurality of reaction channels of a microfluidic chip to identify
DNA sequence variations in each of a plurality of patient samples to diagnose a disease status, a disease risk or predict
a therapeutic drug response for each patient, said method comprising:
(A) for each patient sample, providing a flow of genomic DNA sample within one of the reaction channels, each reaction channel
having a different patient sample;

(B) introducing a plurality of different assay-specific reagent solutions to the flow of genomic DNA sample in a reaction
channel to form a plurality of test mixtures and simultaneously perform different assays on the test mixtures in the reaction
channel, wherein each of the plurality of the test mixtures in the reaction channel undergoes a different assay;

(C) performing a PCR procedure on the test mixture within the reaction channel;
(D) performing a thermal melt procedure on the test mixture after performing the PCR procedure;
(E) collecting data on the thermal melt procedure;
(F) performing an analysis of the collected thermal melt data with a controller programmed to ascertain the presence or absence
of a DNA sequence variation of interest within the test mixture;

(G) performing a conversion of the results of the analyzing step with the controller into a digital signal by designating
a result indicating the presence of the DNA sequence variation of interest as 1 and designating a result indicating the absence
of the DNA sequence variation of interest as 0, or vice versa;

(H) performing an analysis of the outcome of step G with the controller for each of the patient samples to determine whether
the outcome is ambiguous, such that if the controller determines that the outcome is ambiguous, the controller causes the
repeating of steps B through G at least once with the same assay-specific reagent solutions used in step B to improve the
confidence in the determination of the presence or absence of the DNA sequence variation of interest;

(I) performing an analysis of the outcome of step G with the controller for each of the patient samples to determine whether
the outcome is ambiguous, such that if the controller determines that the outcome is not ambiguous, the controller causes
the repeating of steps B through H at least one time with a different assay-specific reagent solution adapted to identify
a different DNA sequence variation relevant to the diagnosis or prediction; and

(J) performing a statistical analysis on the results of steps B through I with the controller to derive a diagnosis or a prediction
and a level of confidence in the diagnosis or prediction.

US Pat. No. 9,766,139

COMPOUND CALIBRATOR FOR THERMAL SENSORS

Canon U.S. Life Sciences,...

1. A method of calibrating at least one of a plurality of thermal control elements in a thermal device, wherein the thermal
device comprises a microfluidic chip having more than one thermal zones, each thermal zone being in thermal contact with at
least one of said plurality of thermal control elements, wherein the microfluidic chip comprises at least one microfluidic
channel that is in thermal contact with each of the more than one thermal zones, the method comprising:
a) introducing a fluid slug comprising a calibrator into the microfluidic channel such that the fluid slug fills the entire
portion of the microfluidic channel that is in thermal contact with a first thermal zone in thermal contact with at least
a first thermal control element, and fills less than the entire portion of the microfluidic channel that is in thermal contact
with a second thermal zone;

b) utilizing at least the first thermal control element to perform a thermal variation within the first thermal zone, generating
a first thermal response profile for said calibrator, wherein the fluid slug is held in place within the first thermal zone
during the calibration of the at least first thermal control element based on detecting an edge position of the fluid slug
in the second thermal zone;

c) identifying at least a first feature of said first thermal response profile and generating a first relation between a known
temperature of said first feature and a first measurement value of said first thermal control element; and

d) calculating one or more calibration coefficients for said at least first thermal control element based on at least said
first relation.

US Pat. No. 9,527,083

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A microfluidic device for performing biological reactions comprising:
a microfluidic chip having a plurality of microfluidic channels and a plurality of multiplexed heater electrodes, wherein
the heater electrodes are part of a multiplex circuit including a common lead connecting the heater electrodes to a power
supply, each of the microfluidic channels having associated therewith one of the heater electrodes; and

a control system configured to regulate power applied to each heater electrode by varying a duty cycle, the control system
being further configured to determine the temperature of each heater electrode by determining the resistance of each heater
electrode.

US Pat. No. 9,061,279

COMPOSITION AND METHOD FOR IN-SYSTEM PRIMING MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. A method of in-system priming a microfluidic device comprising:
(a) applying a priming solution to a microfluidic device having at least one microchannel, wherein the priming solution comprises
a 1×PCR buffer containing a surfactant, wherein a concentration of the surfactant is from a factor of about 5 to a factor
of about 150 of the critical micelle concentration (CMC) of the surfactant; and

(b) applying an external pressure to the microfluidic device to drive or move the priming solution along the at least one
microchannel.

US Pat. No. 9,513,196

METHODS AND SYSTEMS FOR MICROFLUIDIC DNA SAMPLE PREPARATION

Canon U.S. Life Sciences,...

1. A method of purifying DNA in a sample in a microfluidic device comprising the steps of:
(a) mixing the sample and a lysis buffer in a mixing region of a microfluidic device;
(b) selectively lysing the cellular membranes of cells in the sample without lysing the nuclear membranes of cells in a cell
lysing region of the microfluidic device to produce intact nuclei from the cells;

(c) flowing a sample flow containing the intact nuclei into a cell trapping region and trapping the intact nuclei using a
filter in the cell trapping region of the microfluidic device while flowing a cross-flow buffer and other components of the
sample across the sample flow and through the filter into a waste collection region of the microfluidic device;

(d) lysing the intact nuclei trapped by the filter;
(e) releasing the DNA from the lysed nuclei; and
(f) collecting the released DNA in a DNA collection region of the microfluidic device.

US Pat. No. 9,109,961

COMPOUND CALIBRATOR FOR THERMAL SENSORS

Canon U.S. Life Sciences,...

1. A method for calibrating a thermal control element, comprising:
providing a compound calibrator in thermal contact with said thermal control element, wherein the compound calibrator is a
mixture of two or more amplicons of a nucleic acid;

utilizing said thermal control element to perform a thermal variation on said compound calibrator to generate a thermal response
profile for said compound calibrator;

identifying a first feature of said thermal response profile and generating a first relation between a known temperature of
said first feature and a first measurement value of said thermal control element;

identifying a second feature of said thermal response profile and generating a second relation between a known temperature
of said second feature and a second measurement value of said thermal control element;

calculating one or more calibration coefficients for said thermal control element based on said first and second relations.

US Pat. No. 9,861,985

SLUG CONTROL DURING THERMAL CYCLING

Canon U.S. Life Sciences,...

1. A method for controlling the position of a slug in a microfluidic device, comprising:
(a) providing a slug in a microfluidic channel of the microfluidic device;
(b) exciting a fluorescent dye in said slug;
(c) acquiring an image including a portion of the microfluidic channel having the slug emitting the fluorescence and a portion
of the microfluidic channel without the slug;

(d) processing intensity profile data from a region of interest in said image, wherein the region of interest encompasses
a portion of the microfluidic channel without the slug, the intensity profile data including fluorescence intensity presented
as a function of a position along the microfluidic channel;

(f) identifying a position of said slug within said region of interest from said intensity profile data;
(e) controlling the position of said slug within the microfluidic channel by moving the slug to a target position based on
the identified position of said slug in said microfluidic channel; and

(g) repeating steps (b)-(e) at a regular rate while the slug is moving along the microfluidic channel.
US Pat. No. 9,592,510

REAL-TIME PCR IN MICRO-CHANNELS

Canon U.S. Life Sciences,...

1. A method of performing real-time PCR comprising the steps of:
a) moving test solution containing real-time PCR reagents into a channel;
b) uniformly cycling the temperature in a defined section of the channel in order to achieve PCR; and
c) simultaneously measuring the intensity of the fluorescent signal emitted at a plurality of locations along said defined
section of said channel at a specific temperature and time during a PCR temperature cycle, wherein each of the simultaneously
measured intensities of the fluorescent signal emitted at each location along the channel corresponds to the same temperature
and to the same point in time within a PCR cycle.

US Pat. No. 9,542,526

METHOD AND SYSTEM FOR TEMPERATURE CORRECTION IN THERMAL MELT ANALYSIS

Canon U.S. Life Sciences,...

1. A computerized method for correcting a measured melting temperature of a nucleic acid in a sample, wherein the melting
temperature refers to the dissociation of double stranded DNA into single stranded DNA, wherein a first reference sample comprising
a nucleic acid, and a second nucleic acid containing sample are utilized, comprising:
a) determining a ratio of the concentration of the nucleic acid in the first reference sample to the concentration of the
nucleic acid in the second nucleic acid containing sample, wherein the first and second samples contain identical nucleic
acids in different concentrations;

b) using an imaging system and a temperature control system in communication with a computer to measure the melting temperature
of the nucleic acid in the second nucleic acid containing sample (Tm second sample measured) and the melting temperature of the nucleic acid in the first reference sample (Tm1);

c) determining a temperature compensation value from said ratio and determining the change of enthalpy (?H0) of the nucleic acid in the first reference sample,

wherein the temperature compensation value is ?Tm, wherein ?Tm is Tm2?Tm1,

wherein Tm1 is the measured melting temperature of the nucleic acid in the first reference sample and

Tm2 is calculated by the equation


wherein R is the gas constant and C1/C2 is the ratio of the concentration of the nucleic acid in the first reference sample
to the concentration of the nucleic acid in the second nucleic acid containing sample;

wherein the determining a temperature compensation value is performed by the computer; and
d) correcting the measured melting temperature of the nucleic acid in the second nucleic acid containing sample with the temperature
compensation, wherein the correcting the measured melting temperature is performed by the computer based upon the formula
Tm second sample corrected=Tm second sample measured??Tm.

US Pat. No. 9,234,236

MICROFLUIDIC CHIP FEATURES FOR OPTICAL AND THERMAL ISOLATION

Canon U.S. Life Sciences,...

1. A microfluidic device comprising:
at least one microfluidic channel having fluorescent indicators;
wherein the micro fluidic device comprises a reflecting surface, separate from the at least one microfluidic channel, configured
to cause light passing through or emanating from the at least one microfluidic channel having fluorescent indicators toward
said reflecting surface to be redirected by total internal reflection.

US Pat. No. 9,919,314

AIR COOLING SYSTEMS AND METHODS FOR MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. A method for air cooling a microfluidic device having one or more microfluidic channels, one or more inlet ports, one or
more outlet ports and one or more heat sinks, the method comprising:
receiving cooling air through an inlet of a first duct of a bi-level cooling manifold;
using the first duct of the cooling manifold to isolate the cooling air from the one or more inlet ports of the microfluidic
device;

directing the cooling air to a vertical channel of the first duct of the cooling manifold using an upper confinement channel
of the first duct of the cooling manifold;

directing the cooling air to the one or more heat sinks of the microfluidic device using the vertical channel of the first
duct of the cooling manifold;

heating the cooling air using the heat sinks of the microfluidic device;
directing the heated air into a lower confinement channel of a second duct of the cooling manifold using an opening of the
second duct of the cooling manifold;

using the second duct of the cooling manifold to isolate the heated air from the one or more inlet ports of the microfluidic
device; and

directing the heated air to an outlet of the second duct using the lower confinement channel.

US Pat. No. 9,873,122

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A method for determining a selected characteristic of at least one electronic component in a network comprising more than
one of said electronic components to control power supplied to the at least one electronic component, wherein all electronic
components in the network are all either of resistive type, inductive type, or capacitive type, the method comprising the
steps of:
a. generating N distinct partitions of the components, wherein
N is equal to the number of components in the network,
each partition divides the components into at least a source set and a drain set, wherein the source set and the drain set
don't have common components and

at least one of the source set and the drain set in each partition comprises two or more of the components;
b. for each partition:
(1) connecting a source voltage to each component in the source set, wherein the drain set is disconnected from the source
voltage, wherein the source set does not include the source voltage,

(2) connecting a drain voltage to each component in the drain set, and
(3) measuring the equivalent selected characteristic of a circuit between the source voltage and the drain voltage, wherein
the circuit comprises a parallel combination of each component in the source set in series with a parallel combination of
each component in the drain set and the drain voltage is measured at the drain set;

c. computing the selected characteristic of the at least one component based at least in part on the stored equivalent characteristics;
and

d. controlling power supplied to the at least one electronic component in the network based on the computed characteristic
of the at least one component.

US Pat. No. 9,138,744

FLUID INTERFACE CARTRIDGE FOR A MICROFLUIDIC CHIP

Canon U.S. Life Sciences,...

1. A microfluidic assembly comprising:
microfluidic assay chip having a plurality of microfluidic channels, each microfluidic channel having an inlet port for delivering
fluid to a proximal end of the microfluidic channel and an outlet port for removing fluid from a terminal end of the microfluidic
channel;

an interface cartridge having a larger width and length than said microfluidic assay chip, said interface cartridge having
formed therein a plurality of fluid delivery channels in fluid-communication with one or more microfluidic channels in the
microfluidic assay chip, each fluid delivery channel having a fluid delivery port configured to deliver fluid from the associated
fluid delivery channel to an inlet port of one of the microfluidic channels.

US Pat. No. 9,823,135

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A microfluidic device for performing biological reactions, the microfluidic device comprising:
a microfluidic chip having at least one microfluidic channel and at least one heater electrode associated with the microfluidic
channel; and

a control system configured to regulate power applied to the heater electrode by varying a duty cycle, the control system
being further configured to measure a current flowing through the at least one heater electrode indicative of the temperature
of the heater electrode during both the power-on portion of the duty cycle and the power-off portion of the duty cycle.

US Pat. No. 10,137,673

METHODS AND SYSTEMS FOR CONTINUOUS FLOW CELL LYSIS IN A MICROFLUIDIC DEVICE

Canon U.S. Life Sciences,...

1. A method for fabricating a microfluidic device using soft-lithography replica molding, comprising:fabricating a first piece of the microfluidic device in a mold, the first piece including at least one microchannel having features sized to provide for mechanical cell lysis, the features including constricted regions and non-constricted regions separating the constricted regions, the non-constricted regions being shaped to converge at the ends into the constricted regions, wherein fabricating the first piece comprises:
pouring a first layer of a thermoplastic prepolymer onto the mold; and
partially curing the first layer under exposure to UV-light;
fabricating a second piece of the microfluidic device on a glass slide, wherein fabricating the second piece comprises:
coating the glass slide with a layer of the thermoplastic prepolymer; and
partially curing the layer coated on the glass slide under exposure to UV-light;
removing the partially cured first piece from the mold, wherein the partially cured first layer is removed from the mold without loss of the features of the at least one microchannel;
contacting the first piece to the second piece; and
bonding the first piece and the second piece together on the glass slide by curing the contacted first and second pieces under exposure to UV-light.
US Pat. No. 9,983,155

METHOD AND APPARATUS FOR GENERATING THERMAL MELTING CURVES IN A MICROFLUIDIC DEVICE

Canon U.S. Life Sciences,...

1. A system for performing a thermal denaturation analysis of a nucleic acid in a microfluidic device, the system comprising:a microfluidic device having at least one microfluidic channel;
a loading device configured to introduce a fluid comprising the nucleic acid and amplification reagents into the microfluidic channel so that the fluid flows into the channel;
a heating system in communication with a temperature controller configured to cycle a fluid temperature in at least one portion of the microfluidic channel so that the nucleic acid undergoes amplification; and
the temperature controller configured to continuously increase a temperature of the fluid containing the amplified nucleic acid in at least one portion of the channel to cause denaturation of the nucleic acid after the amplification is completed;
a detector configured to measure a detectable property emanating from the fluid containing the amplified nucleic acid in the at least one portion of the microfluidic channel as a function of a continuously increasing temperature, the detectable property being indicative of the extent of denaturation of the nucleic acid, the detectable property decreasing as a nucleic acid molecule denatures in real time, wherein the amplification and the measurement of the detectable property are performed in the same channel, wherein the fluid flow during the amplification is subject to the same flow control as the fluid flow during the thermal melt denaturation; and
a controller configured to generate a thermal property curve representing the measured detectable property as a function of the continuously increasing temperature and determine a nucleic acid melting temperature based on the thermal property curve.

US Pat. No. 9,939,336

SYSTEMS AND METHODS FOR AUTO-CALIBRATION OF RESISTIVE TEMPERATURE SENSORS

Canon U.S. Life Sciences,...

1. A method comprising:providing a source node maintained at a predetermined source voltage;
providing a ground node maintained at a predetermined ground voltage;
providing a bridge circuit comprising:
a first resistance temperature detector connected between the source and a first measurement node,
a first reference resistor connected between the first measurement node and the ground node,
a potentiometer connected between the source node and a reference node, and
a scaling resistor connected between the reference node and the ground node; and
providing a first programmable gain instrumentation amplifier wherein a first input to the first programmable gain instrumentation amplifier is connected to the reference node, a second input to the first programmable gain instrumentation amplifier is connected to the first measurement node, and the voltage output of the first programmable gain instrumentation amplifier is representative of the temperature sensed by the first resistance temperature detector,
modulating a current passing through the first resistance temperature detector by using a bypass circuit connected between the first measurement node and the ground node, controlling the voltage output of the first programmable gain instrumentation amplifier indicative of a temperature of the first resistance temperature detector by:
(a) setting the resistance value of the potentiometer to a first resistance value;
(b) setting the gain of the first programmable gain instrumentation amplifier to a first gain value;
(c) measuring the voltage output from the first programmable gain instrumentation amplifier;
(d) in the case that the measured voltage output is above a predetermined target value, adjusting the resistance value of the potentiometer in a first direction;
(e) in the case that the measured voltage is below the predetermined target value, adjusting the resistance value of the potentiometer in a direction opposite to the first direction; and
(f) repeating steps (c) through (e) until the measured voltage output from the first programmable gain instrumentation amplifier is equal to the predetermined target value.
US Pat. No. 9,701,997

COMPOSITION AND METHOD FOR IN-SYSTEM PRIMING MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. A priming solution comprising:
a 1×PCR buffer containing a surfactant, wherein a concentration of the surfactant is from a factor of about 5 to a factor
of about 150 of the critical micelle concentration (CMC) of the surfactant; and

a dye or a marker at concentration from 5 ?M to about 500 ?M for monitoring the priming solution.
US Pat. No. 9,630,158

METHOD OF DELIVERING PCR SOLUTION TO MICROFLUIDIC PCR CHAMBER

Canon U.S. Life Sciences,...

1. A method of delivering a solution flow, comprising:
causing a reagent and a primer to flow into a first mixing channel;
stopping the reagent and the primer in the first mixing channel by keeping both ends of the first mixing channel at the same
pressure for at least a threshold amount of time so as to allow the reagent and the primer to mix, thereby forming a reagent/primer
mixture;

causing a buffer to flow into a second mixing channel, the first and second mixing channels being located on a mixing chip,
wherein the mixing chip is separate and in fluid communication with a PCR chip through an interface chip, wherein the PCR
chip is configured for performing an amplification reaction;

after holding the reagent and the primer in the first mixing channel for at least the threshold amount of time, drawing, from
the first mixing channel, the reagent/primer mixture into a common exit channel located on the mixing chip; and

adding DNA samples to the reagent/primer mixture while the reagent/primer mixture is in the interface chip.
US Pat. No. 9,903,003

SYSTEMS FOR PROCESSING MULTIPLE ASSAYS BACKGROUND

Canon U.S. Life Sciences,...

44. A microfluidic reaction system comprising:
a microfluidic device including a reaction chip;
a flow controller configured to:
build a blanking slug by drawing a blanking solution into a microfluidic channel of the reaction chip until the leading edge
of the blanking slug reaches a blanking slug building target region; and

build a sample slug by drawing a sample solution into the microfluidic channel of the reaction chip until the trailing edge
of the blanking slug reaches a sample slug building target region;

a heating system including:
a heating element associated with a thermal zone of the reaction chip, wherein the thermal zone is defined as a portion of
the microfluidic channel uniformly heated by the heating element at any point in time; and

a heating controller configured to control the heating element to:
heat a portion of the sample slug in the thermal zone of the reaction chip to cycle the temperature of the portion of the
sample slug in the thermal zone according to a polymerase chain reaction (PCR) amplification profile; and

after completion of the PCR reaction, heat the portion of the sample slug by heating the thermal zone of the reaction chip
according to a temperature ramp profile, wherein the PCR amplification profile and the temperature ramp profile are sequentially
provided for the portion of the sample slug in the thermal zone by the same heating element; and

a detection system configured to, during heating of the portion of the sample slug in the thermal zone according to the temperature
ramp profile, measure fluorescence from the portion of the sample slug in the thermal zone;

wherein the flow controller is additionally configured to draw a second portion of blanking solution into the microfluidic
channel after the detection system measures fluorescence from the portion of the sample slug in the thermal zone, so that
the sample slug is moved from the thermal zone.

US Pat. No. 10,092,902

FLUID INTERFACE CARTRIDGE FOR A MICROFLUIDIC CHIP

Canon U.S. Life Sciences,...

1. An apparatus to increase chip capacity in a microfluidic chip, comprising:a primary microfluidic chip with at least one microfluidic channel enclosed therein and connection ports associated with the at least one microfluidic channel, wherein each microfluidic channel is in fluid communication with a connector channel having two connection ports incorporated into the primary microfluidic chip; and
a secondary cartridge comprising reagent/waste wells in fluid communication with fluidic extension channels and connection holes associated with the fluidic extension channels;
wherein the reagent/waste wells of the secondary cartridge are configured to be removably connected to the connector channel via the connection holes of the fluidic extension channels of the secondary cartridge, each connector channel connecting two connection holes of the fluidic extension channels to the connection ports.
US Pat. No. 9,657,331

COMPOSITION AND METHOD FOR IN-SYSTEM PRIMING MICROFLUIDIC DEVICES

Canon U.S. Life Sciences,...

1. A priming solution comprising:
a 1×PCR buffer containing a surfactant, wherein a concentration of the surfactant is from a factor of about 5 to a factor
of about 150 of the critical micelle concentration (CMC) of the surfactant; and

a dye or a marker at concentration from 5 ?M to about 500 ?M for monitoring the priming solution.
US Pat. No. 9,840,737

METHODS AND SYSTEMS FOR SEQUENTIAL DETERMINATION OF GENETIC MUTATIONS AND/OR VARIANTS

Canon U.S. Life Sciences,...

1. A method of sequentially analyzing a biological sample for the presence of a disease causing mutation comprising the steps
of:
(a) obtaining the biological sample and amplifying at least one exon of a gene of interest in the biological sample;
(b) finding at least one exon having a mutation by screening each amplified exon of the gene of interest in the biological
sample for the presence of a mutation to find each exon having a mutation without genotyping the mutation;

(c) for the at least one exon having a mutation found in step (b), subjecting to amplification in a separate amplification
reaction each location containing a particular mutation within the at least one exon having a mutation by using primers specific
to the particular mutation, each separately amplified section being shorter than the exon; and

(d) separately screening each amplified section within each exon found in step (b) for each particular mutation occurring
in that specific exon to genotype the mutation identified in step (b).

US Pat. No. 9,732,380

SYSTEMS AND METHODS FOR MONITORING THE AMPLIFICATION AND DISSOCIATION BEHAVIOR OF DNA MOLECULES

Canon U.S. Life Sciences,...

1. A DNA analysis system, comprising:
a chip comprising a microchannel for receiving a sample of solution comprising nucleic acid and for providing a path for the
sample to traverse;

an image sensor having a pixel array, wherein at least a portion of the microchannel is within a field of view of the pixel
array; and

an image sensor controller configured to change an area of the pixel array for (a) reading a first area of the pixel array
at a time when the sample is within a field of view of the first area, thereby producing first image data, and for (b) reading
a second area of the pixel array at a time when the sample is within a field of view of the second area, wherein a center
of the first area is spatially separated from a center of the second area, wherein

the image sensor controller is configured to use the first data to determine the size of the second area and the size of the
second area is less than the size of the first area.

US Pat. No. 9,580,744

METHOD AND APPARATUS FOR APPLYING CONTINUOUS FLOW AND UNIFORM TEMPERATURE TO GENERATE THERMAL MELTING CURVES IN A MICROFLUIDIC DEVICE

Canon U.S. Life Sciences,...

1. A method of performing thermal melt analysis of a nucleic acid in a microfluidic device, the method comprising:
providing a microfluidic device having at least one microfluidic channel;
introducing fluid comprising the nucleic acid and reagents into the at least one microfluidic channel;
continuously flowing the fluid through the at least one microfluidic channel while continuously and at a constant rate increasing
the temperature of the entire fluid stream as it moves through the at least one microfluidic channel by uniformly heating
the entire fluid stream;

measuring at two or more points along the microfluidic channel, while continuously flowing the fluid through the at least
one microfluidic channel, a detectable property emanating from the fluid, wherein the detectable property indicates an extent
of denaturation of the nucleic acid as the temperature of the entire fluid stream is increasing; and

generating a melt curve of the nucleic acid, each point of the melt curve based upon combined measurements of the detectable
property performed at two or more points along the microfluidic channel.

US Pat. No. 9,540,686

SYSTEMS AND METHODS FOR THE AMPLIFICATION OF DNA

Canon U.S. Life Sciences,...

1. A system for amplifying DNA, comprising:
a microfluidic chip comprising a sample channel and a heat exchange channel formed within the microfluidic chip and sufficiently
close to the sample channel such that a heat exchange fluid in the heat exchange channel can cause a sample in the sample
channel to gain or lose heat at desired levels, wherein the heat exchange channel is configured to exchange heat with two
sides of the sample channel;

a first reservoir having an output port coupled to an input of the heat exchange channel through a first forward valve and
having an input port coupled to an output of the heat exchange channel through a first return valve, said first reservoir
storing a first heat exchange fluid;

a second reservoir having an output port coupled to the input of the heat exchange channel through a second forward valve
and having an input port coupled to the output of the heat exchange channel through a second return valve, said second reservoir
storing a second heat exchange fluid;

a third reservoir having an output port coupled to the input of the heat exchange channel through a third forward valve and
having an input port coupled to the output of the heat exchange channel through a third return valve, said third reservoir
storing a third heat exchange fluid, wherein each of the first, second, and third reservoirs is divided into two chambers
fluidly connected with each other by a valve, the fluid being released into the heat exchange channel from a first chamber
and returned back from the heat exchange channel into the second chamber;

a temperature control system configured to: (a) regulate the heat exchange fluid stored in the first reservoir at a first
temperature, (b) regulate the heat exchange fluid stored in the second reservoir at a second temperature, and (c) regulate
the heat exchange fluid stored in the third reservoir at a third temperature;

one or more pumps;
an imaging system including an excitation source and an image capturing device configured to image a biological reaction within
the sample channel through a sample channel region unobstructed by the heat exchange channel; and

a controller configured to operate said valves and said one or more pumps such that:
(a) for a first period of time, the first heat exchange fluid stored in the first reservoir enters the heat exchange channel,
but the second and third heat exchange fluids stored in the second and third reservoirs, respectively, do not enter the heat
exchange channel;

(b) for a second period of time, the second heat exchange fluid stored in the second reservoir enters the heat exchange channel,
but the first and third heat exchange fluids stored in the first and third reservoirs, respectively, do not enter the heat
exchange channel; and

(c) for a third period of time, the third heat exchange fluid stored in the third reservoir enters the heat exchange channel,
but the first and second heat exchange fluids stored in the first and second reservoirs, respectively, do not enter the heat
exchange channel, wherein

the first period of time is different than the second period of time, which is different than the third period of time, and
the first temperature is different than the second temperature, which is different than the third temperature, wherein the
first heat exchange fluid is returned back to the first reservoir prior to directing the second heat exchange fluid to the
heat exchange channel and the second heat exchange fluid is returned back to the second reservoir prior to directing the third
heat exchange fluid to the heat exchange channel.

US Pat. No. 9,777,318

SYSTEMS AND METHODS FOR MONITORING THE AMPLIFICATION OF DNA

Canon U.S. Life Sciences,...

1. A method of performing real-time PCR comprising:
moving a sample of test solution containing real-time PCR reagents through a channel;
while the sample is moving through a section of the channel:
cycling the temperature of the sample in order to achieve PCR;
illuminating the sample with excitation light; and
using a fiber optic probe to capture fluorescent light emitted from the sample,
wherein said fiber optic probe includes: (i) an optical fiber having a proximal end and a distal end, and (ii) a probe head
connected to and positioned adjacent to the distal end of the optical fiber, said probe head positioned such that at least
a portion of the section of the channel is within a field of view of the probe head; and

measuring the intensity of said fluorescent light.

US Pat. No. 10,066,977

MICROFLUIDIC FLOW MONITORING

Canon U.S. Life Sciences,...

1. A system for monitoring a flow rate of a solution in a microfluidic channel comprising:(a) a chip comprising the microfluidic channel;
(b) a fluid movement system for moving a fluid through the channel;
(c) a flow marker introduction system for introducing a flow marker into the channel, wherein said flow marker introduction system comprises (i) a piezoelectric nozzle, a bubble jet head, or a sipper; (ii) a converging inlet channel comprising a valve, and (iii) a directed energy source;
(d) an illumination system for illuminating the flow marker at two measuring points, wherein there is no illumination of the flow marker between the two measuring points;
(e) a detection system configured for measuring a transit time during which the flow marker moves within the channel over a distance between the two measuring points by detecting signals caused by the flow marker flowing past the two illuminated measuring points and taking a time stamp in response to the flow marker flowing past each of the two illuminated measuring points, wherein the flow marker can be detected at the two illuminated measuring points; and
(f) a flow calculation system for determining flow marker velocity based on the transit time, and correlating the flow marker velocity to the flow rate.

US Pat. No. 9,724,695

SYSTEMS AND METHODS FOR AMPLIFYING NUCLEIC ACIDS

Canon U.S. Life Sciences,...

32. An adaptable apparatus for performing a thermocyclic process comprising:
a microfluidic chip comprising a first material and having a fluid channel formed therein; and
one or more thermal distribution elements comprising a second material; wherein the one or more thermal distribution elements
are placed in thermal communication with an associated portion of said microfluidic chip, each said thermal distribution element
being constructed and arranged to distribute thermal energy from an external thermal energy source, including a heater, substantially
uniformly over said associated portion of said microfluidic chip, wherein each of the one or more thermal distribution elements
is positioned between and in contact with the microfluidic chip and the heater,

wherein the size and positioning of the one or more heaters and thermal distribution elements in relation to the fluid channel
creates two or more adaptable temperature zones on the microfluidic chip, thereby defining said associated portion as one
of said temperature zones within said microfluidic chip,

a detector configured to detect emissions originating from one or more locations within the channel, wherein the one or more
heaters and thermal distribution elements positioned on the same side as the detector relative to the microfluidic chip obstruct
optical communication between the detector and a fluid in the fluid channel passing through temperature zones in thermal communication
with the thermal distribution elements, wherein a fluid passing through temperature zones not in thermal communication with
the thermal distribution elements is in optical communication with the detector,

wherein the size and positioning of the one or more heaters and thermal distribution elements in relation to the microfluidic
chip is changeable,

wherein said channel is arranged such that the fluid flowing through the channel would enter and exit each of said temperature
zones of the microfluidic chip a plurality of times.

US Pat. No. 9,829,389

MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES

Canon U.S. Life Sciences,...

1. A system comprising:
a multiplex circuit comprising a plurality of multiplexed heater electrodes sharing a common lead connecting the electrodes
to a power supply;

a controller having instruction for:
sequentially measuring the combined series resistances for N number of distinct sensor pairs, such that each of the plurality
of sensors is included in at least one of the measured sensor pairs, wherein N is the number of sensors in the plurality of
sensors, and the measured sensor pairings are such that the individual resistance of each of the plurality of sensors may
be determined; and

determining by a processor the individual resistance of at least one of the plurality of sensors based upon the measured combined
series resistances.

US Pat. No. 9,938,519

METHODS AND SYSTEMS FOR DNA ISOLATION ON A MICROFLUIDIC DEVICE

Canon U.S. Life Sciences,...

1. A method of isolating DNA from cells in a sample comprising the steps of:(a) selectively lysing the cellular membranes of the cells in the sample without lysing the nuclear membranes of the cells to produce intact nuclei from the cells;
(b) separating the intact nuclei from the sample by a nuclei size exclusion barrier in a nuclei separation region of a microfluidic device;
(c) resuspending the separated nuclei in an elution buffer in the nuclei separation region of the microfluidic device;
(d) delivering the resuspended nuclei to a nuclei lysis region of a microfluidic device, wherein the nuclei lysis region is spatially separated from the nuclei separation region; and
(e) lysing the resuspended nuclei in the nuclei lysis region of the microfluidic device to release and isolate the DNA.

US Pat. No. 9,962,692

METHODS, DEVICES, AND SYSTEMS FOR FLUID MIXING AND CHIP INTERFACE

Canon U.S. Life Sciences,...

1. A method for delivering a reaction mixture to a microfluidic chip comprising a docking receptacle, an access tube and a reservoir, the method comprising:engaging a pipette tip which has docking feature and which contains the reaction mixture, with a reservoir of the microfluidic chip by engaging the docking receptacle of the reservoir with the docking feature of the pipette tip;
producing a bead of the reaction mixture on the exterior of the pipette tip, wherein the bead makes contact with the access tube of the microfluidic chip;
pulling at least a first portion of the reaction mixture from the bead into the access tube of the microfluidic chip while the bead is attached to the pipette tip, wherein the pipette tip comprises a disk attached to a proximal end of the pipette tip to provide additional surface area for the bead to attach; and
removing the tip of the pipette from the microfluidic device, wherein a second portion of the reaction mixture remains in the bead externally attached to the pipette tip as it is removed from contact with the access tube of the microfluidic chip, leaving reaction mixture only inside the access tube and not in the reservoir of the microfluidic chip.

US Pat. No. 9,987,627

METHOD AND MOLECULAR DIAGNOSTIC DEVICE FOR DETECTION, ANALYSIS AND IDENTIFICATION OF GENOMIC DNA

Canon U.S. Life Sciences,...

1. A microfluidic system comprising:a cartridge configured to isolate genomic material comprising:
a reaction chamber;
an ejector head;
a waste outlet;
wherein the reaction chamber is connected to the ejector head and with the waste outlet by a first channel and second channel, respectively;
at least one binding and release substrate, wherein the at least one binding and release substrate lies within the reaction chamber and is configured to bind and release at least a portion of a sample comprising genomic material and undesirable material;
two electrodes, wherein a first electrode is positioned in the reaction chamber and a second electrode is positioned in the first channel connecting the reaction chamber and the ejector head; and
a power source to effect changes in a state of the binding and release substrate;
the genomic material releasably bound to the binding and release substrate, wherein the genomic material is released from the at least one binding and release substrate based upon a change in the state of the binding and release substrate;
wherein the two electrodes are constructed and arranged to direct charged genomic material into the ejector head while waste material is directed into the waste outlet;
and
a microfluidic chip, separate from the cartridge, comprising at least one microfluidic inline reaction channel, wherein the genomic material is ejected from the ejector head into an inlet port of the microfluidic chip across an airspace between the cartridge and the microfluidic chip.

US Pat. No. 9,964,557

APPARATUS FOR OPTICAL MICROFLUIDICS SLUG EDGE DETECTION

Canon U.S. Life Sciences,...

1. A system comprising:a channel having two adjacent fluid slugs positioned along the channel, the slugs having a color gradient across an edge;
an image sensor configured to acquire an image of at least a portion of the adjacent fluid slugs in the channel, the acquired image characterized by an intensity signal changing at least along the channel;
non-transitory media in communication with a processing unit, the non-transitory media comprising an edge detection unit in communication with the image sensor, the edge detection unit providing instructions to the processing unit to:
sequentially apply at least two different segmentations to the length of the channel, each segmentation partitioning the length of the channel into two segments, left segment and right segment, at a specific location;
for each of the at least two segmentations, calculate an edge score function proportional to a between class variance for intensity signal values associated with the two adjacent segments, wherein the between class variance is calculated based upon a probability for each of the two adjacent segments and a mean value of the intensity signal in each of the two adjacent segments; and
select a segmentation defining an edge between the two adjacent fluid slugs based at least in part on the edge score function, wherein a location of the edge between the two adjacent slugs is defined as a coordinate of the specific location selected along the length of the channel to separate the two adjacent segments in the selected segmentation.
US Pat. No. 9,573,132

SYSTEM AND METHODS FOR MONITORING THE AMPLIFICATION AND DISSOCIATION BEHAVIOR OF DNA MOLECULES

Canon U.S. Life Sciences,...

1. A method, comprising:
(a) introducing into a microchannel at least one bolus containing nucleic acid;
(b) forcing the bolus to move through the microchannel;
(c) while the bolus is moving through the microchannel, (i) causing a thermal generating apparatus to cycle the temperature
of the bolus to amplify nucleic acid contained in the bolus, wherein the thermal generating apparatus sequentially controls
temperature during amplification and thermal denaturation, the thermal denaturation being performed after the amplification
is completed and (ii) at least once per temperature cycle determining whether a predetermined emission intensity threshold
has been met, wherein the step of determining whether the predetermined emission intensity threshold has been met comprises
using an image sensor to capture an image of the bolus and processing the captured image data to determine the intensity of
light emitted from the bolus;

(d) in response to determining that the predetermined emission intensity threshold has not been met, then repeating step (c);
and

(e) in response to determining that the predetermined emission intensity threshold has been met, then, causing said thermal
generating apparatus to cease the temperature cycling and causing said thermal generating apparatus to gradually increase
the temperature of the bolus, while the bolus is still moving through the microchannel, causing thermal denaturation of dsDNA
within the bolus to transition to ssDNA and using the image sensor to capture images of the bolus.

US Pat. No. 10,376,891

SYSTEM AND METHOD FOR TEMPERATURE REFERENCING FOR MELT CURVE DATA COLLECTION

Canon U.S. Life Sciences,...

1. A system for performing melt curve data collection comprising:a container comprising at least three chambers, wherein at least two of the chambers are spatially separated from each other, and contain a temperature reference material, and at least one chamber contains a DNA test sample;
a heating system to heat the container,
a detection system, wherein a detectable signal from the temperature reference material provides feedback to the heating system; and
a control system for determining a spatial temperature gradient due to temporal fluctuation within the container to calculate a temperature at a location of the DNA test sample based on temperature results from the temperature reference material that surrounds the location of the DNA test sample.

US Pat. No. 10,363,558

SYSTEM AND METHOD FOR SERIAL PROCESSING OF MULTIPLE NUCLEIC ACID ASSAYS

Canon U.S. Life Sciences,...

1. A system for serial processing of nucleic acid assays, comprising:a microfluidic cartridge comprising an interface chip having at least one inlet port and microfluidic channel and a separate reaction chip coupled to the interface chip, the reaction chip having at least one microfluidic channel ending in a T-junction, the T-junction in fluid communication with an associated microfluidic channel of the interface chip;
a flow control module comprising a first pumping system configured to selectively apply pneumatic pressures to the at least one microfluidic channel of the interface chip and a second pumping system to apply a pneumatic pressure to control assay fluid flow through the reaction chip;
the flow control module being configured to:
(a) draw a first fluid into the microfluidic channel of the interface chip via the inlet port of the interface chip and stop the first fluid in the interface chip when the T-junction is loaded with the first fluid;
(b) create a first fluid segment in the microfluidic channel of the reaction chip by drawing the first fluid from the associated microfluidic channel of the interface chip into the microfluidic channel of the reaction chip;
(c) draw a second fluid into the microfluidic channel of the interface chip via the inlet port of the interface chip while maintaining the position of the first fluid segment in the reaction chip and stop the second fluid in the interface chip when the T-junction is loaded with the second fluid; and
(d) create a second fluid segment in the microfluidic channel of the reaction chip by drawing the second fluid from the associated microfluidic channel of the interface chip into the microfluidic channel of the reaction chip;
a temperature measurement and control system configured to control and measure a temperature of one or more portions of the at least one microfluidic channel of the reaction chip; and
a fluorescence imaging system comprising one or more illumination elements and an imaging sensor and configured to create images of fluorescent emissions from materials within the at least one microfluidic channel of the reaction chip.

US Pat. No. 10,266,873

OPTICAL SYSTEM FOR HIGH RESOLUTION THERMAL MELT DETECTION

Canon U.S. Life Sciences,...

1. A system comprising:a microfluidic chip comprising at least one microchannel;
a sensor element configured to generate a storable image of at least a portion of the microchannel, wherein the image is generated of a predetermined non-uniformly heated region of interest (ROI) including ROIs heated by calibrated and non-calibrated heating elements, respectively, wherein the sensor element is configured to read out one or more predetermined portions of the image's pixel array as a sub-ROI;
a controller configured to calibrate fluorescence data acquired from the sub-ROIs heated by non-calibrated heating elements with fluorescence data acquired from the sub-ROIs heated by calibrated heating elements;
and
a plurality of illumination elements disposed with respect to the sensor element and configured to illuminate a portion of the at least one microchannel to be imaged by the sensor element,
wherein at least one of the illumination elements comprises an illumination assembly comprising:
an LED;
a mask disposed in front of the LED and having an opening formed therein so as to control an area illuminated by the illumination assembly;
a filter along an optic path of the illumination assembly for controlling the spectral content of light emitted by the illumination assembly, and
a lens for imaging an area with light emitted by the illumination assembly,
wherein the LED, the mask, the filter, and the lens are aligned along an optic axis of the illumination assembly.

US Pat. No. 10,226,772

COMBINED THERMAL DEVICES FOR THERMAL CYCLING

Canon U.S. Life Sciences,...

1. A nucleic acid amplification system comprising:a microfluidic device comprising at least one microfluidic channel,
means to flow a fluid sample through the at least one microfluidic channel,
a first heating device to heat and cool the fluid sample to achieve nucleic acid amplification,
a second heating device to heat and cool the fluid sample to achieve nucleic acid amplification,
a first temperature sensing device to sense the temperature of the fluid sample,
a second temperature sensing device to sense the temperature of the fluid sample, and
a controller to energize and deenergize the first and second heating devices based on readings provided by the first and second temperature sensing device, wherein the first and second heating devices have different temperature resolutions.

US Pat. No. 10,386,288

SYSTEM AND METHOD OF LABEL-FREE CYTOMETRY BASED ON BRILLOUIN LIGHT SCATTERING

Canon U.S. Life Sciences,...

1. A method for simultaneously obtaining one or more metrics associated with a Brillouin scattering spectrum at multiple points within a sample, the method comprising:illuminating the sample by a light beam from an illumination source along a first direction;
collecting by one or more lenses a Brillouin scattered light emitted from the sample in response to the illuminating light beam;
sending the Brillouin scattered light to an optical arrangement to induce a spectral dispersion and to a detection unit to generate a spatio-spectral pattern of the Brillouin scattered light, wherein the optical arrangement and the detection unit are positioned along a second direction;
detecting the spatio-spectral pattern of the Brillouin scattering light onto the detection unit, wherein multiple points of the sample along the illuminating light beam are measured simultaneously;
calibrating the spectral pattern at each spatial point at the detection unit;
calculating the one or more Brillouin metrics at each measured sample point based on the detected spatio-spectral pattern; and
using the calculated one or more Brillouin metrics from the multiple points of the sample to identify one or more properties of the sample.

US Pat. No. 10,359,353

SYSTEM AND METHOD OF LABEL-FREE CYTOMETRY BASED ON BRILLOUIN LIGHT SCATTERING

Canon U.S. Life Sciences,...

1. A method for simultaneously obtaining one or more metrics associated with a Brillouin scattering spectrum at multiple points within a sample, the method comprising:illuminating the sample by a light beam from an illumination source along a first direction;
collecting by one or more lenses a Brillouin scattered light emitted from the sample in response to the illuminating light beam;
sending the Brillouin scattered light to an optical arrangement to induce a spectral dispersion and to a detection unit to generate a spatio-spectral pattern of the Brillouin scattered light, wherein the optical arrangement and the detection unit are positioned along a second direction;
detecting the spatio-spectral pattern of the Brillouin scattering light onto the detection unit, wherein multiple points of the sample along the illuminating light beam are measured simultaneously;
calibrating the spectral pattern at each spatial point at the detection unit;
calculating the one or more Brillouin metrics at each measured sample point based on the detected spatio-spectral pattern; and
using the calculated one or more Brillouin metrics from the multiple points of the sample to identify one or more properties of the sample.