# US 10,767,219 B2 · Disposable, integrated microfluidic cartridge and methods of making and using same

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- Number: US 10,767,219 B2
- Document ID: US10767219B2
- Status: Granted
- Kind: B2
- Application number: 15/125,124
- Filed: 2015-03-11
- Published: 2017-01-19
- Granted: 2020-09-08
- Earliest claimed: 2014-03-11
- Assignees: Illumina, Inc.
- Applicants: Illumina, Inc.
- Inventors: Poorya Sabounchi; Behnam Javanmardi; Tarun Khurana; Philip Paik; Yan-You Lin
- Web page: https://patentgrep.com/patent/US10767219B2

## CPC

- C12Q 1/6869: Methods for sequencing
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- B01L 3/502707: {characterised by the manufacture of the container or its components}
- B01L 3/502738: {characterised by integrated valves (throttle valves in microfluidic sample containers B01L3/502746)}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2200/12: Specific details about manufacturing devices
- B01L 2300/0636: Integrated biosensor, microarrays
- B01L 2300/0654: Lenses; Optical fibres
- B01L 2300/0809: rectangular shaped
- B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution
- B01L 2300/0874: Three dimensional network
- B01L 2300/0887: Laminated structure
- B01L 2300/123: Flexible; Elastomeric
- B01L 2300/165: Specific details about hydrophobic, oleophobic surfaces
- B01L 2300/1827: using resistive heater
- B01L 2400/0487: fluid pressure, pneumatics
- B01L 2400/0644: rotary valves
- B01L 2400/0655: pinch valves
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

The disclosed embodiments concern microfluidic cartridges for detecting biological reactions. In some embodiments, the microfluidic cartridges are configured to perform sequencing operations on a nucleic acid sample. In one aspect, a microfluidic cartridge includes a stack of fluidics layers defining channels and valves for processing the nucleic acid sample to be sequenced, and a solid state CMOS biosensor integrated in the stack. The biosensor has an active area configured to detect signals of biological reactions, wherein substantially all of the active area is available for reagent delivery and illumination during operation. In another aspect, a microfluidic cartridge includes: (a) a flow cell including a reaction site area encompassing one or more reaction sites; (b) fluidics channels for delivering reactants to and/or removing reactants from the reaction site area; (c) a biosensor having an active area configured to detect signals of biological reactions in the reaction site area. The reaction site area is proximal to the active area of the biosensor and the reaction site area spans substantially all of the active area of the biosensor. In some embodiments, the fluidics channels do not substantially overlap with the active area of the biosensor. Methods for manufacturing and operating the microfluidic cartridges are also disclosed.

## Claims

### Claim 1

A microfluidic cartridge for detecting biological reactions, comprising:
(a) a flow cell including a reaction site area encompassing one or more reaction sites; (b) fluidics channels for delivering reactants to and/or removing reactants from the reaction site area; and (c) an image sensor having an active area configured to detect signals of biological reactions in the reaction site area,
wherein:
the reaction site area is proximal to the active area of the image sensor;
the reaction site area spans substantially all of the active area of the image sensor;
the fluidics channels do not substantially overlap with the active area of the image sensor; the flow cell comprises a sequencing chamber; the sequencing chamber is formed on a sequencing chamber layer; the image sensor is disposed in an opening of a sequencing chamber bottom layer under the sequencing chamber layer; and the fluidics channels are formed on a fluidics channels layer under the sequencing chamber bottom layer.

### Claim 2 (depends on 1)

The microfluidic cartridge of claim 1, wherein
the microfluidic cartridge is configured to perform sequencing operations on a nucleic acid sample, and
the detected signals of biological reactions are indicative of nucleotide base types involved in the biological reactions.

### Claim 3 (depends on 2)

The microfluidic cartridge of claim 2, wherein the flow cell comprises a substrate of hydrophilic regions for nucleic acid attachment and amplification surrounded by hydrophobic regions.

### Claim 4 (depends on 1)

The microfluidic cartridge of claim 1, wherein the reaction site area spans all of the active area of the image sensor.

### Claim 5 (depends on 1)

The microfluidic cartridge of claim 1, wherein the fluidics channels do not overlap with the active area of the image sensor.

### Claim 6 (depends on 1)

The microfluidic cartridge of claim 1, wherein the image sensor comprises a photo detector.

### Claim 7 (depends on 6)

The microfluidic cartridge of claim 6, wherein the photo detector comprises a CMOS/CCD sensor.

### Claim 8 (depends on 7)

The microfluidic cartridge of claim 7, wherein the CMOS sensor is about 9200 μm long, about 8000 μm wide, and about 800-1000 μm thick.

### Claim 9 (depends on 1)

The microfluidic cartridge of claim 1, further comprising: a PCR region, a reagent mixing and distributing region, and one or more membrane valves that are configured to reversibly stop the PCR region from fluidic communication with the reagent mixing and distribution region or the flow cell including the reaction site area.

### Claim 10 (depends on 9)

The microfluidic cartridge of claim 9, wherein the PCR region comprises a plurality of PCR channels.

### Claim 11 (depends on 9)

The microfluidic cartridge of claim 9, wherein the reagent mixing and distributing region comprises a plurality of reagent channels and/or reagent supplies.

### Claim 12 (depends on 9)

The microfluidic cartridge of claim 9, further comprising a rotary valve that is configured to fluidly connect the PCR region to the reagent mixing and distributing region.

### Claim 13 (depends on 12)

The microfluidic cartridge of claim 12, wherein the rotary valve is further configured to fluidly connect the reagent mixing and distributing region to the flow cell including a reaction site area.