# US 2023/0058091 A1 · Solid reagent containment unit, in particular for a transportable microfluidic device for sample preparation and molecule analysis

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- Number: US 2023/0058091 A1
- Document ID: US20230058091A1
- Status: Published application
- Kind: A1
- Application number: 17/981,299
- Filed: 2022-11-04
- Published: 2023-02-23
- Earliest claimed: 2017-12-28
- Assignees: STMicroelectronics S.r.l.
- Applicants: STMICROELECTRONICS S.r.l.
- Inventors: Marco CEREDA; Lillo RAIA; Alessandro Paolo BRAMANTI
- Web page: https://patentgrep.com/patent/US20230058091A1

## CPC

- B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules}
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/502738: {characterised by integrated valves (throttle valves in microfluidic sample containers B01L3/502746)}
- B01L 3/502753: {characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation}
- B01L 3/5029: {using swabs}
- B01L 3/508: {Rigid containers without fluid transport within}
- B01L 3/5082: {Test tubes per se}
- B01L 3/52: {Containers specially adapted for storing or dispensing a reagent (B01L3/02 takes precedence; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002)}
- B01L 3/527: {for a plurality of reagents}
- B01L 9/527: {for microfluidic devices, e.g. used for lab-on-a-chip}
- B29C 43/14: in several steps
- B29C 43/18: incorporating preformed parts or layers, e.g. compression moulding around inserts or for coating articles {(B29C43/206 takes precedence)}
- C12N 15/1003: {Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor}
- B01L 3/5023: {with a sample being transported to, and subsequently stored in an absorbent for analysis}
- B01L 3/502707: {characterised by the manufacture of the container or its components}
- B01L 3/50825: {Closing or opening means, corks, bungs}
- B01L 3/523: {with means for closing or opening}
- B01L 2200/025: Align devices or objects to ensure defined positions relative to each other
- B01L 2200/026: Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L 2200/0621: Control of the sequence of chambers filled or emptied
- B01L 2200/0631: Purification arrangements, e.g. solid phase extraction [SPE]
- B01L 2200/0642: Filling fluids into wells by specific techniques
- B01L 2200/0668: Trapping microscopic beads
- B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid
- B01L 2200/0689: Sealing
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2200/141: Preventing contamination, tampering
- B01L 2200/16: Reagents, handling or storing thereof
- B01L 2300/022: Transponder chips
- B01L 2300/023: Sending and receiving of information, e.g. using Bluetooth®
- B01L 2300/044: pierceable, e.g. films, membranes
- B01L 2300/0609: Holders integrated in container to position an object
- B01L 2300/0681: Filter
- B01L 2300/08: Geometry, shape and general structure
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0819: Microarrays; Biochips
- B01L 2300/0829: Multi-well plates; Microtitration plates
- B01L 2300/0848: Specific forms of parts of containers
- B01L 2300/0851: Bottom walls
- B01L 2300/0858: Side walls
- B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
- B01L 2300/087: Multiple sequential chambers
- B01L 2300/12: Specific details about materials
- B01L 2300/16: Surface properties and coatings
- B01L 2300/1827: using resistive heater
- B01L 2400/04: Moving fluids with specific forces or mechanical means
- B01L 2400/0406: capillary forces
- B01L 2400/0421: electrophoretic flow
- B01L 2400/043: magnetic forces
- B01L 2400/0487: fluid pressure, pneumatics
- B01L 2400/049: vacuum
- B01L 2400/0677: phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
- B29C 43/021: {characterised by the shape of the surface}
- B29L 2031/753: {Medical equipment; Accessories therefor (bloodbags, medical bags B29L2031/7148; artificial eyes B29L2011/0008)}
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B29C: SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING (making preforms B29B11/00; making laminated products by combining previously unconnected layers which become one product whose layers will remain together B32B37/00 - B32B41/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- B29L: INDEXING SCHEME ASSOCIATED WITH SUBCLASS B29C, RELATING TO PARTICULAR ARTICLES

## Abstract

A solid reagent containment unit is formed by a support; a frame body fixed to the support and delimiting internally, together with the support, an analysis volume; a reagent-adhesion structure within the analysis volume; and at least one reagent cavity, which extends within the reagent-adhesion structure. The reagent-adhesion structure is of an adhesion material embossable at temperatures lower by 6-8° C. than its own melting point and has a melting point such as not to interfere with the analysis. The reagent cavity forms a retention wall, laterally surrounding the reagent cavity, and houses dried reagents. The adhesion material is chosen among wax, such as paraffin, a polymer, such as polycaprolactone, a solid fat, such as cocoa butter, and a gel, such as hydrogel or organogel.

## Claims

### Claim 1

A method for sample preparation and molecule analysis, comprising:
introducing a portable microfluidic device into a support;
introducing a sample into a sample inlet opening of the portable microfluidic device;
extracting molecules from the sample to obtain an analyzable sample;
transferring the analyzable sample into an analysis volume of a solid reagent containment unit of the portable microfluidic device;
re-hydrating a solid reagent in the solid reagent containment unit with the analyzable sample; and
conducting an analysis on the analyzable sample, wherein the portable microfluidic device includes:
an extraction chamber in fluidic connection with an opening for introducing samples;
a waste chamber extending along the extraction chamber;
a collector extending along the extraction chamber and the waste chamber;
a fluidic circuit connecting the extraction chamber, the waste chamber, and the collector; and
the solid reagent containment unit which includes:
a support;
a frame body fixed to the support, the frame body delimiting inside, together with the support, an analysis volume;
a reagent-adhesion structure within the analysis volume; and
a reagent cavity, which extends within the reagent adhesion structure, wherein:
the reagent adhesion structure is formed by an inert adhesion material embossable at lower temperatures than a melting point of the adhesion material,
the reagent cavity houses solid reagents, and
the collector comprises an analysis recess designed to house the solid reagent containment unit, with the analysis recess in fluidic connection with the reagent cavity.

### Claim 2 (depends on 1)

The method for sample preparation and molecule analysis of claim 1, wherein the support of the solid reagent containment unit is an integrated circuit chip.

### Claim 3 (depends on 1)

The method for sample preparation and molecule analysis of claim 1, wherein integrated circuit chip includes a heater that is aligned with the reagent cavity of the solid reagent containment unit

### Claim 4 (depends on 1)

The method of claim 1, wherein:
transferring the sample into the analysis volume of the solid reagent containment unit of the portable microfluidic device further includes:
opening a valve to open a fluidic pathway to the analysis chamber along which the analyzable sample travels along to the analysis chamber; and
generating suction pressure with a pump in fluid communication with the transportable microfluidic device;
re-hydrating the solid reagent in the solid reagent containment unit with the analyzable sample further includes introducing the analyzable sample to the reagent cavity containing the solid reagent.

### Claim 5 (depends on 1)

The method of claim 1, wherein extracting molecules from the sample to obtain the analyzable sample includes:
introducing a preparation reagent into the extraction chamber in which the sample and magnetic beads are present by activating an light emitting diode (LED) opening a one-shot valve along an inlet fluidic recess of the transportable microfluidic device, by opening a first valve hole through deforming a first shutter, and by generating a suction pressure with a pump;
activating an anchor actuator moving and rotating an anchor in the extraction chamber to stir and mix the preparation reagent, the sample, and the magnetic beads in the extraction chamber extracting the molecules from the sample and adhering the magnetic beads to the molecules;
removing the preparation reagents from the extraction chamber while leaving the magnetic beads and the molecules within the extraction chamber by opening a second valve hole through deforming a second shutter and activating a blocking actuator attracting the magnetic beads and holding the magnetic beads to which the molecules are adhered within the extraction chamber;
flushing the molecules by introducing in at least one flushing liquid into the extraction chamber; and
forming the analyzable sample by introducing an elution liquid purifying the molecules and separating the molecules from the magnetic beads.

### Claim 6 (depends on 5)

The method of claim 5, wherein removing the preparation reagent from the extraction chamber includes introducing the preparation reagent to the waste chamber within the transportable microfluidic device delimited by the first wall, the second wall, and the body.

### Claim 7 (depends on 5)

The method of claim 5, wherein flushing the molecules by introducing the at least one flushing liquid into the extraction chamber further includes introducing a plurality of flushing liquids in succession to the extraction chamber.

### Claim 8 (depends on 1)

The method of claim 1, wherein the molecules are a nucleic acid present within the sample.

### Claim 9

A method, comprising:
introducing a sample into an extraction chamber of a portable microfluidic device by introducing the sample into a sample inlet opening of the portable microfluidic device, the extraction chamber being defined by a first wall of the portable microfluidic device, a second wall of the portable microfluidic device, and a body of the portable microfluidic device that is between the first wall and the second wall;
extracting molecules from the sample in the extraction chamber obtaining an analyzable sample;
transferring the analyzable sample to an analysis chamber of the portable microfluidic device delimited by the body and the first wall;
exposing the analyzable sample to a solid reagent in a reagent cavity of a reagent unit and on an integrated circuit chip of the reagent unit, the reagent cavity being delimited by a sidewall of an embossable holding material of the reagent unit and a surface of the integrated circuit chip; and
analyzing the analyzable sample exposed to the solid reagent within the reagent cavity.

### Claim 10 (depends on 9)

The method of claim 9, wherein extracting molecules from the sample in the extraction chamber obtaining the analyzable sample includes:
introducing a preparation reagent into the extraction chamber in which the sample and magnetic beads are present by activating an light emitting diode (LED) opening a one-shot valve along an inlet fluidic recess of the portable microfluidic device, by opening a first valve hole through deforming a first shutter, and by generating a suction pressure with a pump;
activating an anchor actuator moving and rotating an anchor in the extraction chamber to stir and mix the preparation reagent, the sample, and the magnetic beads in the extraction chamber extracting the molecules from the sample and adhering the magnetic beads to the molecules;
removing the preparation reagents from the extraction chamber while leaving the magnetic beads and the molecules within the extraction chamber by opening a second valve hole through deforming a second shutter and activating a blocking actuator attracting the magnetic beads and holding the magnetic beads to which the molecules are adhered within the extraction chamber;
flushing the molecules by introducing at least one flushing liquid into the extraction chamber; and
forming the analyzable sample by introducing an elution liquid purifying the molecules and separating the molecules from the magnetic beads.

### Claim 11 (depends on 10)

The method of claim 10, wherein transferring the analyzable sample to the analysis chamber of the portable microfluidic device delimited by the body and the first wall further includes opening a third valve hole along a fluidic pathway that extends from the extraction chamber to the analysis chamber by deforming a third shutter and generating the suction pressure with the pump.

### Claim 12 (depends on 10)

The method of claim 10, wherein removing the preparation reagent from the extraction chamber includes discharging the preparation reagent from the portable microfluidic device through a fluidic outlet of the portable microfluidic device.

### Claim 13 (depends on 10)

The method of claim 10, wherein removing the preparation reagent includes introducing the preparation reagent to a waster chamber within the portable microfluidic device delimited by the first wall, the second wall, and the body.

### Claim 14 (depends on 10)

The method of claim 10, further comprising heating the solid reagent with a heater of the integrated circuit aligned with the reagent cavity in which the solid reagent is present.

### Claim 15

A method, comprising:
introducing a sample into an extraction chamber of a portable microfluidic device dissolving a tablet present within the extraction chamber releasing a plurality of magnetic beads within the extraction chamber, the extraction chamber being defined by a first wall of the portable microfluidic device, a second wall of the portable microfluidic device, and a body of the portable microfluidic device that is between the first wall and the second wall;
extracting molecules from the sample in the extraction chamber obtaining an analyzable sample including:
introducing preparation reagents within the extraction chamber in which the sample is present;
activating an anchor actuator to rotate an anchor within the extraction chamber to stir and mix the sample with the preparation reagents extracting the molecules from the sample and adhering the molecules to the magnetic beads;
magnetically attracting the magnetic beads towards a blocking actuator by activating the blocking actuator to; and
removing the preparation reagent spent from extracting the molecules from the sample;
transferring the analyzable sample to an analysis chamber of the portable microfluidic device delimited by the body and the first wall;
exposing the analyzable sample to a solid reagent in a reagent cavity of a reagent unit, the reagent cavity being delimited by a sidewall of an embossable holding material of the reagent unit and a surface of an integrated circuit chip of the reagent unit; and
analyzing the analyzable sample exposed to the solid reagent within the reagent cavity.

### Claim 16 (depends on 15)

The method of claim 15, wherein extracting the molecules from the extraction chamber obtaining the analyzable sample further includes:
introducing a preparation reagent into the extraction chamber in which the sample and magnetic beads are present by activating an light emitting diode (LED) opening a one-shot valve along an inlet fluidic recess of the portable microfluidic device, by opening a first valve hole through deforming a first shutter, and by generating a suction pressure with a pump;
flushing the molecules by introducing at least one flushing liquid into the extraction chamber; and
forming the analyzable sample by introducing an elution liquid purifying the molecules and separating the molecules from the magnetic beads.

### Claim 17 (depends on 16)

The method of claim 16, wherein flushing the molecules by introducing the at least one flushing liquid into the extraction chamber further includes introducing respective liquids of a plurality of flushing liquids into the extraction chamber.

### Claim 18 (depends on 17)

The method of claim 17, wherein introducing the respective liquids of the plurality of flushing liquids into the extraction chamber further includes introducing successively the respective liquids of the plurality of flushing liquids into the extraction chamber

### Claim 19 (depends on 18)

The method of claim 18, wherein the respective liquids of the plurality of flushing liquids are stored within a plurality of reagent chambers within the portable microfluidic device, and the plurality of reagent chambers are delimited by the first wall, the second wall, and the body.

### Claim 20 (depends on 18)

The method of claim 18, wherein the molecules are a nucleic acid present within the sample.