US20040223874A1 - Biochemical reaction cartridge - Google Patents

Biochemical reaction cartridge Download PDF

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Publication number
US20040223874A1
US20040223874A1 US10/811,917 US81191704A US2004223874A1 US 20040223874 A1 US20040223874 A1 US 20040223874A1 US 81191704 A US81191704 A US 81191704A US 2004223874 A1 US2004223874 A1 US 2004223874A1
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United States
Prior art keywords
chamber
cartridge
liquid
passage
specimen
Prior art date
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Abandoned
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US10/811,917
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English (en)
Inventor
Yasuyuki Numajiri
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Canon Inc
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Canon Inc
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Priority claimed from JP2003097136A external-priority patent/JP4111505B2/ja
Application filed by Canon Inc filed Critical Canon Inc
Assigned to CANON KABUSHIKI KAISHA reassignment CANON KABUSHIKI KAISHA ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: NUMAJIRI, YASUYUKI
Publication of US20040223874A1 publication Critical patent/US20040223874A1/en
Priority to US11/549,204 priority Critical patent/US7988913B2/en
Abandoned legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures 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
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures 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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
    • F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
    • F04B3/00—Machines or pumps with pistons coacting within one cylinder, e.g. multi-stage
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures 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
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures 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 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
    • F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
    • F04B53/00—Component parts, details or accessories not provided for in, or of interest apart from, groups F04B1/00 - F04B23/00 or F04B39/00 - F04B47/00
    • F04B53/10—Valves; Arrangement of valves
    • F04B53/102—Disc valves
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
    • F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
    • F04B53/00—Component parts, details or accessories not provided for in, or of interest apart from, groups F04B1/00 - F04B23/00 or F04B39/00 - F04B47/00
    • F04B53/16—Casings; Cylinders; Cylinder liners or heads; Fluid connections
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/087—Multiple sequential chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1822—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using Peltier elements
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples

Definitions

  • the present invention relates to a technology to analyze cell, microorganism, chromosome, nuclei acid, etc., in a specimen by utilizing a biochemical reaction. More specifically, the present invention relates to a biochemical reaction cartridge for use in the analysis and a biochemical treatment apparatus for effecting the biochemical reaction in the cartridge.
  • analyzers for analyzing specimens uses an immunological procedure utilizing antigen-antibody reaction or a procedure utilizing nuclei acid hybridization.
  • protein or single-stranded nucleic acid such as antibody or antigen, which specifically connects with a material or substance to be detected, is used as a probe and is fixed on a surface of solid phase, such as fine particles, beads or a glass plate, thus effecting antigen-antibody reaction or nuclei acid hybridization.
  • an antigen-antibody compound or double-stranded nucleic acid is detected by a labeled antigen or labeled nucleic acid, which causes a specific interaction such that a labeled material having a high detection sensitivity, such as an enzyme, a fluorescent material or a luminescent material, is supported, thus effecting detection of presence or absence of the material to be detected or quantitative determination the detected material.
  • a labeled material having a high detection sensitivity such as an enzyme, a fluorescent material or a luminescent material
  • U.S. Pat. No. 5,445,934 has disclosed a so-called DNA (deoxyribonucleic acid) array wherein a large number of DNA probes having mutually different base sequences are arranged on a substrate in array form.
  • Anal. Biochem., 270(1), pp. 103-111 (1999) has disclosed a process for preparing a protein-array, like the DNA array, such that various species of proteins are arranged on a membrane filter. By using these DNA and protein arrays and the like, it has become possible to effect a test on a large number of items at the same time.
  • JP-A Japanese Laid-Open Patent Application
  • Japanese Laid-Open Patent Application (JP-A) (Tokuhyo) Hei 11-509094 has disclosed a biochemical reaction cartridge, including DNA array, in which a plurality of chambers are disposed and a solution is moved by a differential pressure so as to permit a reaction such as extraction, amplification or hybridization of DNA in a specimen within the cartridge.
  • JP-A Japanese Laid-Open Patent Application
  • 5,690,763 has disclosed a constitution for reacting a three-dimensionally curved passage through sheet lamination
  • U.S. Pat. Nos. 6,167,910 and 6,494,230 have disclosed structures of ⁇ -TAS (micro-total analysis system) wherein a passage is provided between a first layer and a second layer and between a second layer and a third layer, constituting a three-layer structure, and the respective passages are partially connected with each other.
  • ⁇ -TAS micro-total analysis system
  • a disposable cartridge containing a necessary solution is used from the viewpoints of prevention of secondary infection or contamination and usability but the cartridge containing a pump is expensive.
  • An object of the present invention is to provide a disposable biochemical reaction cartridge having a structure capable of causing a sequence of a biochemical reaction to proceed by moving a solution under the action of an external pump without containing a pump and capable of preventing the solution from flowing out of the cartridge.
  • Another object of the present invention is to provide a biochemical treatment apparatus for effecting the biochemical reaction within the cartridge by using the biochemical reaction cartridge described above.
  • Another object of the present invention is to provide a method o fusing a biochemical reaction cartridge capable of ensuring appropriate movement in such a manner that in a biochemical reaction cartridge for effecting movement of liquid therein, an optimum passage is selected and used properly with respect to movement of a reagent or a specimen only requiring injection into a subsequent chamber and movement of a reaction liquid requiring reciprocating motion.
  • a biochemical reaction cartridge comprising:
  • a second chamber for containing therein a reagent which contributes to a biochemical reaction
  • the plurality of nozzle ports communicate with the first or second chamber, and fluid is present between the plurality of nozzle ports and the first or second chamber and is pressurized or depressurized by the plurality of nozzles to move the specimen and/or the reagent and/or the reaction liquid, thereby to effect a sequence of a biochemical reaction within the cartridge.
  • a biochemical treatment apparatus comprising:
  • a cartridge mounting portion for mounting a cartridge having a plurality of chambers containing a solution for biochemically treating a specimen
  • control means for controlling a fluid pressure in the cartridge through the nozzle portions
  • control means controls the fluid pressure so that the solution in the cartridge is moved only in the cartridge.
  • a biochemical treatment process for effecting biochemical treatment in a cartridge having a plurality of chambers containing a solution for biochemically treating a specimen comprising:
  • a biochemical reaction cartridge comprising:
  • a bottom position of a first connecting portion for connecting the first chamber to the first passage is higher than a bottom position of a second connecting portion for connecting the first chamber to the second passage.
  • FIG. 1 is a perspective view of an embodiment of the biochemical reaction cartridge according to the present invention.
  • FIG. 2 is a plan view of the biochemical reaction cartridge.
  • FIG. 3 is a block diagram of a treatment apparatus for controlling movement of liquid and various reactions within the biochemical reaction cartridge.
  • FIG. 4 is a flow chart of a treatment procedure.
  • FIG. 5 is a longitudinal sectional view of a part of a chamber.
  • FIG. 6 is a longitudinal sectional view of another part of the chamber.
  • FIG. 7 is a longitudinal sectional view of another part of the chamber.
  • FIG. 8 is a longitudinal sectional view of a part of a chamber according to another embodiment.
  • FIG. 1 is an external view of a biochemical reaction cartridge 1 in this embodiment.
  • a specimen port 2 for injecting a specimen such as blood by a syringe (injector) or the like is disposed and sealed up with a rubber cap.
  • a rubber cap is fixed on each of the nozzle ports 3 .
  • the other side surface of the cartridge 1 has a similar structure.
  • a body of the biochemical reaction cartridge 1 comprises transparent or semitransparent synthetic resin, such as polymethyl methacrylate (PMMA), acrylonitrile-butadiene-styrene (ABS) copolymer, polystyrene, polycarbonate, polyester or polyvinyl chloride.
  • PMMA polymethyl methacrylate
  • ABS acrylonitrile-butadiene-styrene copolymer
  • polystyrene polycarbonate
  • polyester or polyvinyl chloride polyester or polyvinyl chloride
  • FIG. 2 is a plan view of the biochemical reaction cartridge 1 .
  • 10 nozzle ports 3 a to 3 j are provided and also on the other side surface thereof, 10 nozzle ports 3 k to 3 t are provided.
  • the respective nozzle ports 3 a to 3 t communicate with chambers 5 , which are portions or sites for storing the solution or causing a reaction, through corresponding air passages 4 a to 4 t , respectively.
  • the nozzle ports 3 n , 3 p , 3 q and 3 s are not used, these nozzle ports do not communicate with the chambers 5 and are used as reserve ports. More specifically, in this embodiment, the nozzle ports 3 a to 3 j communicate with the chambers 5 a to 5 j through the passages 4 a to 4 j , respectively.
  • the nozzle ports 3 k , 3 l , 3 m , 3 o , 3 r and 3 t communicate with the chambers 5 k , 5 l , 5 m , 5 o , 5 r and 5 t through the passages 4 k , 4 l , 4 m , 4 o , 4 r and 4 t , respectively.
  • the specimen port 2 communicates with a chamber 7 .
  • the chambers 5 a , 5 b , 5 c and 5 k communicate with the chamber 7
  • the chambers 5 g and 5 o communicate with a chamber 8
  • the chambers 5 h , 5 i , 5 j , 5 r and 5 t communicate with a chamber 9 .
  • the chamber 7 communicate with the chamber 8 via a passage 10
  • the chamber 8 communicates with the chamber 9 via a passage 11 .
  • the chambers 5 d , 5 e , 5 f , 5 l and 5 m communicate via passages 6 d , 6 e , 6 f , 6 l and 6 m , respectively.
  • a square hole is provided.
  • a DNA microarray 12 on which several tens to several hundreds of thousand of different species of DNA probes are arranged in high density on a surface of solid phase, such as a glass plate having a size of ca. square centimeter, with the probe surfaces up, is attached.
  • the DNA probes are regularly arranged in a matrix form, and an address (position determined by the number of row and the number of column on the matrix) of each of the DNA probes is readily read as information.
  • the genes to be tested includes, e.g., genetic polymorphism of each individual in addition to infections viruses, bacteria and disease-associated genes.
  • a first hemolytic agent containing EDTA (ethylenediaminetetraacetic acid) for destructing cell wall and a second hemolytic agent containing a protein modifying agent such as a surfactant are stored, respectively.
  • An eluent, comprising a buffer of low-concentration salt, for eluting DNA from the magnetic particles is stored in the chamber 5 d , a mixture liquid for PCR (polymeraze chain reaction) comprising a primer, polymerase, a dNTP (deoxyribonucleotide triphosphate), a buffer, Cy-3dUTP containing a fluorescent agent, etc., is stored in the chamber 5 g .
  • a cleaning agent containing a surfactant for cleaning a fluorescence-labeled specimen DNA, which is not subjected to hybridization, and a fluorescence label is stored in the chamber 5 i .
  • the chamber 5 e is a chamber in which dust other than DNA of blood accumulates
  • the chamber 5 f is a chamber in which waste of the first and second extraction cleaning liquids in the chambers 5 l and 5 m accumulate
  • the chamber 5 r is a chamber in which waste liquid of the first and second cleaning liquids accumulate
  • the chambers 5 k , 5 o and 5 t are blank chambers provided for preventing the solution to flow into the nozzle ports.
  • FIG. 3 is a schematic view of the treatment apparatus for controlling movement of the solution within the biochemical reaction cartridge and various reactions.
  • the biochemical reaction cartridge 1 is mounted on a table 13. Further, on the table 13, an electromagent 14 to be actuated at the time of extracting DNA or the like from the specimen in the cartridge 1 , a Peltier element 15 for effecting temperature control at the time of amplifying DNA from the specimen through a method such as PCR (polymerase chain reaction), and a Peltier element 16 for effecting temperature control at the time of performing hybridization between the amplified specimen DNA and the DNA probe on the DNA microarray within the cartridge 1 and at the time of cleaning or washing the specimen DNA which is not hybridized, are disposed and connected to a control unit 17 for controlling the entire treatment apparatus.
  • PCR polymerase chain reaction
  • an electric (motor-driven) syringe pumps 18 and 19 and pump blocks 22 and 23 each of which is a port for discharging or sucking in air by these pumps 18 and 19 and is provided with 10 pump nozzles 20 or 21 on its side surface, are disposed.
  • a plurality of electric switching (selector) valves are disposed and connected to the control unit 17 together with the pumps 18 and 19 .
  • the control unit 17 is connected to an input unit 24 to which inputting by a tester is performed.
  • the control unit 17 controls the pump nozzles 20 and 21 so that each of the respective 10 pump nozzles is selectively opened and closed with respect to the electric syringe pumps 18 and 19 , respectively.
  • the tester when the tester injects blood as a specimen into the cartridge 1 through the rubber cap of the specimen port 2 by a syringe or an injector, the blood flows into the chamber 7 . Thereafter, the tester places the biochemical reaction cartridge 1 on the table 13 and moves the pump blocks 22 and 23 i directions of arrows indicated in FIG. 3 by operating an unshown lever, whereby the pump nozzles 20 and 21 are injected into the cartridge 1 through the corresponding nozzle ports 3 at the both side surfaces of the cartridge 1 .
  • the nozzle ports 3 a to 3 t are concentrated at two surfaces, i.e., both side surfaces, of the biochemical reaction cartridge 1 , so that it is possible to simplify shapes and arrangements of the electric syringe pumps 18 and 19 , the electric switching valves, the pump blocks 22 and 23 containing the pump nozzles, etc. Further, by effecting such a simple operation that the cartridge 1 is sandwiched between the pump blocks 22 and 23 at the same time while ensuring necessary chambers 5 and passages, it is possible to inject the pump nozzles 20 and 21 and simplify the structure of the pump blocks 22 and 23 .
  • all the nozzle ports 3 a to 3 t are disposed at an identical level, i.e., are arranged linearly, whereby all the heights of the passages 4 a to 4 t connected to the nozzle ports 3 a to 3 t become equal to each other. As a result, preparation of the passages 4 a to 4 t becomes easy.
  • FIG. 4 is a flow chart for explaining a treatment procedure in the treatment apparatus in this embodiment.
  • a step S 1 the control unit 24 opens only the nozzle ports 3 a and 3 b , and air is discharged form the electric syringe pump 18 and sucked in the cartridge 1 from the electric syringe pump 19 , whereby the first hemolytic agent 1 is injected from the chamber 5 a into the chamber 7 containing blood.
  • the solution can flow smoothly without causing splash or scattering thereof at its leading end although it depends on a viscosity of the hemolytic agent and a resistance of the passage.
  • the solution can be caused to flow further smoothly by effecting such a control that a degree of suction of air is linearly increased from the initiation of air discharge from the pump 18 . This is true in the case of subsequent liquid movement.
  • the air supply control can be readily realized by using the electric syringe pumps 18 and 19 . More specifically, after only the nozzle ports 3 a and 3 o are opened, discharge and suction of air are repeated alternately by the pumps 18 and 19 to cause repetitive flow and flowback of the solution of the chamber 7 in the passage 10 , thus stirring the solution. Alternatively, the solution can be stirred while continuously discharging air from the pump 19 to generate bubbles.
  • FIG. 5 is a sectional view of the biochemical reaction cartridge 1 shown in FIG. 2 along a cross section intersecting the chambers 5 a , 7 and 5 k , and shows such a state that the nozzle port 3 a is pressurized by injecting therein the pump nozzle 20 and the nozzle port 3 k is reduced in pressure by injecting therein the pump nozzle 21 , whereby the first hemolytic agent in the chamber 5 a flows into the chamber 7 through the passage 6 a .
  • FIG. 5 in order to clarify a height (level) relationship, a cross section of the passage 10 is also shown.
  • a volume of the first hemolytic agent in the chamber 5 a is determined so that it ensures a requirement. Further, dimensions and positions of the chambers 5 a and 7 are determined so that the liquid level in the chamber 7 is lower than a height (vertical position) of a bottom surface 25 of a connecting portion between the passage 6 a and the chamber 7 when the first hemolytic agent flows into the chamber 7 .
  • a step S 2 in a step S 2 , only the nozzle ports 3 b and 3 k are opened and the second hemolytic agent in the chamber 5 b is caused to flow into the chamber 7 in the same manner as in the case of the first hemolytic agent. Similarly, in a step 53 , the magnetic particles in the chamber 5 c are caused to flow into the chamber 7 .
  • stirring is performed in the same manner as in the step S 1 .
  • step S 3 DNA resulting from dissolution of cells in the steps S 1 and S 2 attaches to the magnetic particles.
  • Cross sectional shapes of the chambers 5 b and 5 c and the passages 6 b and 6 c are the same as those of the chamber 5 a and the passage 6 a . Volumes of the second hemolytic agent and the magnetic particle solution are determined so that they ensure their requirements. Further, dimensions and positions of the chambers 5 b , 5 c and 7 are determined, similarly as in the step S 1 , so that the liquid level in the chamber 7 is lower than height of bottom surfaces of connecting portions between the passages 6 b and 6 c and the chamber 7 .
  • the biochemical reaction cartridge 1 is prepared through ultrasonic fusion bonding of three injection molded parts 1 A, 1 B and 1 C defined by chain double-dashed lines indicated in FIG. 5.
  • the passages 6 a , 6 b and 6 c are identical in height (vertical position) to each other. Accordingly, the associated connection portions are also at the same height.
  • the chambers having the same height as the chambers 5 a , 5 b and 5 c are the chamber 5 k shown in FIG. 1 and the chambers 5 g and 5 o shown in FIG. 2.
  • the reagent is caused to flow from a higher position than the chamber to be moved, so that it is possible to smoothly move reliably the entire amount of the reagent stored in the storage chamber with less resistance. Further, there is such a case that avoidance of generation of bubbles is desired with respect to some reagents. In such case, when the movement of the reagent is performed as described above, the entire amount of the solution can be moved with a simple structure while avoiding the generation of bubbles without monitoring completion of movement of the solution.
  • an electromagnet 14 is turned on and only the nozzle ports 3 e and 3 k are opened. Then, air is discharged from the electric syringe pump 19 and sucked in form the pump 18 to move the solution from the chamber 7 to the chamber 5 e . At the time of movement, the magnetic particles and DNA are trapped in the passage 10 on the electromagnet 14 . The suction and discharge by the pumps 18 and 19 are alternately repeated to reciprocate the solution two times between the chambers 7 and 5 e , whereby a trapping efficiency of DNA is improved. The trapping efficiency can be further improved by increasing the number of reciprocation. In this case, however, it takes a longer treating time by that much.
  • DNA is trapped in a flowing state on such a small passage having a width of about 1-2 mm and a height of about 0.2-1 mm by utilizing the magnetic particles, so that DNA can be trapped with high efficiency. This is also true for RNA and protein.
  • FIG. 6 is a sectional view of the cartridge 1 shown in FIG. 2 along a cross section intersecting the chambers 5 e , 7 and 5 k , and shows a height relationship between the chambers 5 e and 7 and the passage 6 e .
  • the passage 6 e connects the bottom portions of the chambers 5 e and 7 , so that the movement direction of the solution is changed to an opposite direction when the suction by the pump 18 and the discharge by the pump 19 are inverted. As a result, when the suction and the discharge is alternately repeated, it is possible to reciprocate the solution any number of times between the chambers 7 and 5 e.
  • a step S 5 the electromagnet 14 is turned off, and only the nozzle ports 3 f and 3 l are opened. Thereafter, air is discharged from the electric syringe pump 19 and sucked in from the pump 18 to move the first extraction cleaning liquid from the chamber 5 l to the chamber 5 f . At this time, the magnetic particles and DNA trapped in the step S 4 are moved together with the extraction cleaning liquid, whereby cleaning is performed.
  • the electromagnet 14 is turned on, and the reciprocation of two times is similarly performed to recover the magnetic particles and DNA in the passage 10 on the electromagnet 14 and return the solution to the chamber 5 l.
  • a step S 6 cleaning is further performed in the same manner as in the step S 5 by using the second extraction cleaning liquid in the chamber 5 m in combination with the nozzle ports 3 f and 3 m.
  • a step 7 only the nozzle ports 3 d and 3 o are opened while the electromagnet 14 is kept on, and air is discharged from the pump 18 and sucked in from the pump 19 , whereby the eluent in the chamber 5 d is moved to the chamber 8 .
  • the magnetic particles and DNA are separated by the action of the eluent, so that only the DNA is moved together with the eluent to the chamber 8 , and the magnetic particles remain in the passage 10 .
  • extraction and purification of the DNA are performed.
  • the chamber containing the extraction cleaning liquid and the chamber containing waste liquid after the cleaning are separately provided, so that it becomes possible to effect extraction and purification of the DNA in the biochemical reaction cartridge 1 .
  • a step S 8 only the nozzle ports 3 g and 3 o are opened, and air is discharged from the electric syringe pump 18 and sucked in from the pump 19 to cause the PCR agent in the chamber 5 g to flow into the chamber 8 . Further, only the nozzle ports 3 g and 3 t are opened, and air discharge and suction by the pumps 18 and 19 are repeated alternately to cause the solution in the chamber 8 to flow. Thereafter, the returning operation is repeated to effect stirring. Then, the Peltier element 15 is controlled to retain the solution in the chamber 8 at 96° C. for 10 min. Thereafter, a cycle of heating at 96° C./10 sec, 55° C./10 sec, and 72° C./1 min. is repeated 30 times, thus subjecting the eluted DNA to PCR to amplify the DNA.
  • a step S 9 only the nozzle ports 3 g and 3 t are opened, and air is discharged from the electric syringe pump 18 and sucked in from the pump 19 to move the solution in the chamber 8 to the chamber 9 . Further, by controlling the Peltier element 16 , the solution in the chamber 9 is kept at 45° C. for 2 hours to effect hybridization. At this time, discharge and suction of air by the pumps 18 and 19 are repeated alternately to move the solution in the chamber 9 to he passage 6 t . Thereafter, the hybridization proceeds while effecting stirring by repeating the returning operation.
  • a step S 10 while keeping the temperature at 45° C., only the nozzle ports 3 h and 3 r are opened, and air is discharged from the electric syringe pump 18 and sucked in from the pump 19 to cause the first cleaning liquid in the chamber 5 h to flow into the chamber 5 r through the chamber 9 while moving the solution in the chamber 9 to the chamber 5 r .
  • the suction and discharge by the pumps 18 and 19 are repeated alternately to reciprocate the solution two times between the chambers 5 h , 9 and 5 r and finally return the solution to the chamber 5 h .
  • the fluorescence-labeled specimen DNA and the fluorescence label which are not hybridized are cleaned.
  • FIG. 7 is a sectional view of the biochemical reaction cartridge 1 shown in FIG. 2 along a cross section intersecting the chambers 5 h , 9 and 5 r .
  • the cartridge 1 is pressurized by injecting the pump nozzle 20 into the nozzle port 3 h and is reduced in pressure by injecting the pump nozzle 21 into the nozzle port 3 r .
  • FIG. 7 illustrates such a state that the first cleaning liquid is caused to flow into the chamber 5 r through the chamber 9 .
  • a step S 11 while keeping the temperature at 45° C., the cleaning is further effected in the same manner as in the step S 10 by using the second cleaning liquid in the chamber 5 j in combination with the nozzle ports 3 j and 3 r , and the solution is finally returned to the chamber 5 j .
  • the chambers 5 h and 5 j containing the cleaning liquids and the chamber 5 r containing waste liquid after the cleaning are separately provided, so that it becomes possible to effect extraction and purification of the DNA microarray 12 in the biochemical reaction cartridge 1 .
  • a step 12 only the nozzle ports 3 i and 3 r are opened, and air is discharged from the electric syringe pump 18 and sucked in from the pump 19 to move alcohol in the chamber 5 i to the chamber 5 r through the chamber 9 . Thereafter, only the nozzle port 3 i and 3 t are opened, and air is discharged from the pump 18 and sucked in from the pump 19 to dry the chamber 9 .
  • the tester When the tester operates a lever (not shown), the pump blocks 22 and 23 are moved away from the biochemical reaction cartridge 1 . As a result, the pump nozzles 20 and 21 are removed from the nozzle ports 3 of the cartridge 1 . Then, the tester mounts the cartridge 1 in a reader for DNA array, such a known scanner to effect measurement and analysis.
  • FIG. 8 is a sectional view of a biochemical reaction cartridge 1 of this embodiment, and illustrates a cross section intersecting the chambers 5 a , 7 and 5 k shown in FIG. 2 of Embodiment 1. Further, FIGS. 1 to 4 and 7 in Embodiment 1 are also applicable to this embodiment.
  • the biochemical reaction cartridge 1 is pressurized by injecting the pump nozzle 20 into the nozzle port 3 a and reduced in pressure by injecting the pump nozzle 21 into the nozzle port 3 k .
  • FIG. 8 illustrates such a state that a first hemolytic agent in the chamber 5 a is caused to flow into the chamber 7 containing blood through the passage 6 a .
  • a cross section of the passage is also indicated.
  • the passage connecting the chambers 5 a and 7 extends in not only a horizontal direction but also a vertical direction, so that a (vertical) height of a bottom surface 25 of the connection portion between the passage 6 a and the chamber 7 is increased, i.e., a permissible liquid level is increased.
  • a mount of a solution to be contained in the chamber 7 is made larger. If it is not necessary to increase the solution amount, the height of the biochemical reaction cartridge 1 can be decreased.
  • the vertical portion of the passage 6 a is required in this embodiment.
  • it can be provided by using two injection molded parts A and B defined a chain double-dashed line shown in FIG. 8.
  • the passage 6 a may be tilted to have an oblique surface.
  • the movement from the storage chamber is performed with respect to the reagent but may also be performed with respect to liquid specimen or cleaning liquid.
  • the movement of liquid is performed by utilizing pressure application and reduction of air but may also be performed in other manners such that the cartridge 1 is opened at one side surface and only pressurized or reduced in pressure at the other side surface, that a pump which directly moves a solution to be moved is used, and that electrical movement or movement by utilizing a magnetic force is adopted.
  • a predetermined amount of the solution is stored in the storage chamber and all the amount of the solution is moved but, the amount of the moving solution may also be controlled by a liquid amount sensor or a flow rate sensor.
  • the biochemical reaction cartridge according to the present invention moves the solution only therein by an external pump without incorporating a pump to cause an necessary reaction to proceed, so that it becomes possible to provide a disposable cartridge which does not cause outflow of the solution therefrom with an inexpensive structure. As a result, possibilities of secondary infection and contamination are eliminated. Further, the cartridge incorporates therein the necessary solution, so that it is not necessary to prepare a reagent and cleaning liquids. As a result, it becomes possible to realize elimination of labor and prevent an error in selection of the reagent.
  • air pressure within the cartridge is controlled by the (external) pump on the treatment apparatus side to move the solution only within the cartridge, thus causing a necessary biochemical reaction. Accordingly, it becomes possible to effect the biochemical reaction within the cartridge by using the inexpensive biochemical reaction cartridge.
  • the biochemical reaction cartridge according to the present invention can effect movement with reliability and simple structure by properly using an optimum passage with respect to both of movement, for a reagent or specimen, which can be performed only by causing the reagent or specimen to flow into a subsequent chamber, and movement of a reaction liquid requiring reciprocating motion. Further, such an effect that it is possible to move most efficiently a liquid, such as a reagent or an liquid specimen, to a subsequent chamber without causing generation of bubbles, can be attained.

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  • Dispersion Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Engineering & Computer Science (AREA)
  • Mechanical Engineering (AREA)
  • Physics & Mathematics (AREA)
  • Fluid Mechanics (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
US10/811,917 2003-03-31 2004-03-30 Biochemical reaction cartridge Abandoned US20040223874A1 (en)

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JP2003097136A JP4111505B2 (ja) 2003-03-31 2003-03-31 生化学処理装置及び生化学処理方法
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US20120094366A1 (en) * 2006-11-28 2012-04-19 Ludwig Lester F Reconfigurable chemical process systems
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US10073107B2 (en) * 2005-05-25 2018-09-11 Boehringer Ingelheim Vetmedica Gmbh System for operating a system for the integrated and automated analysis of DNA or protein
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EP2661485A4 (fr) * 2011-01-06 2018-11-21 Meso Scale Technologies, LLC Cartouches d'essai et leurs procédés d'utilisation
US20200150142A1 (en) * 2017-06-06 2020-05-14 The Regents Of The University Of California Systems and methods for rapid generation of droplet libraries
US10744502B2 (en) 2016-10-07 2020-08-18 Boehringer Ingelheim Vetmedica Gmbh Analysis device and method for testing a sample
US10816563B2 (en) 2005-05-25 2020-10-27 Boehringer Ingelheim Vetmedica Gmbh System for operating a system for the integrated and automated analysis of DNA or protein
US10953403B2 (en) 2016-10-07 2021-03-23 Boehringer Ingelheim Vetmedica Gmbh Method and analysis system for testing a sample

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US20060013726A1 (en) * 2004-07-14 2006-01-19 Katsumi Munenaka Biochemical reaction cartridge
US8831783B2 (en) 2004-12-13 2014-09-09 Canon Kabushiki Kaisha Biochemical processing apparatus
US20060127277A1 (en) * 2004-12-13 2006-06-15 Canon Kabushiki Kaisha Biochemical processing apparatus
US8025853B2 (en) * 2004-12-13 2011-09-27 Canon Kabushiki Kaisha Biochemical processing apparatus
US9752182B2 (en) 2004-12-23 2017-09-05 Abbott Point Of Care Inc. Molecular diagnostics system and methods
US20100291666A1 (en) * 2004-12-23 2010-11-18 Gordon Bruce Collier Molecular diagnostics system and methods
US8883487B2 (en) 2004-12-23 2014-11-11 Abbott Point Of Care Inc. Molecular diagnostics system and methods
US20160017407A1 (en) * 2005-05-04 2016-01-21 Immunotrex Biologics Inc. Methods for Microorganism Detection and Identification
US10184946B2 (en) * 2005-05-25 2019-01-22 Boehringer Ingelheim Vetmedica Gmbh Method for operating a system for the integrated and automated analysis of DNA or protein
US10073107B2 (en) * 2005-05-25 2018-09-11 Boehringer Ingelheim Vetmedica Gmbh System for operating a system for the integrated and automated analysis of DNA or protein
US10816563B2 (en) 2005-05-25 2020-10-27 Boehringer Ingelheim Vetmedica Gmbh System for operating a system for the integrated and automated analysis of DNA or protein
US8591813B2 (en) 2005-08-15 2013-11-26 Canon Kabushiki Kaisha Reaction cartridge, reaction apparatus and method of moving solution in reaction cartridge
US20070036679A1 (en) * 2005-08-15 2007-02-15 Canon Kabushiki Kaisha Reaction cartridge, reaction apparatus and method of moving solution in reaction cartridge
US10449540B2 (en) * 2006-11-28 2019-10-22 Nri R&D Patent Licensing, Llc General-purpose reconfigurable conduit and reaction chamber microfluidic arrangements for lab-on-chip and miniature chemical processing
US20120094366A1 (en) * 2006-11-28 2012-04-19 Ludwig Lester F Reconfigurable chemical process systems
US20170225163A1 (en) * 2006-11-28 2017-08-10 Lester F. Ludwig General-purpose reconfigurable conduit and reaction chamber microfluidic arrangements for lab-on-chip and miniature chemical processing
US9636655B2 (en) * 2006-11-28 2017-05-02 Lester F. Ludwig Software-reconfigurable conduit and reaction chamber microfluidic arrangements for lab-on-a-chip and miniature chemical processing techologies
EP3109618A3 (fr) * 2009-12-07 2017-03-15 Meso Scale Technologies, LLC. Cartouches de test comprenant une chambre d'échantillon
US9646133B2 (en) * 2010-06-09 2017-05-09 Lester F. Ludwig Computer system and microfluidic instrumentation for next-generation biological signaling network research and applications
US20170235872A1 (en) * 2010-06-09 2017-08-17 Lester F. Ludwig Computer system and microfluidic instrumentation for next-generation biological signaling network research and drug discovery
US20110307182A1 (en) * 2010-06-09 2011-12-15 Ludwig Lester F Computer system and microfluidic instrumentation for next-generation biological signaling network research and applications
US11896978B2 (en) 2011-01-06 2024-02-13 Meso Scale Technologies, Llc. Assay cartridges and methods of using the same
US10814327B2 (en) 2011-01-06 2020-10-27 Meso Scale Technologies, Llc. Assay cartridges and methods of using the same
EP2661485A4 (fr) * 2011-01-06 2018-11-21 Meso Scale Technologies, LLC Cartouches d'essai et leurs procédés d'utilisation
US20140220173A1 (en) * 2011-09-02 2014-08-07 The Regents Of The University Of California Universal hardware platform and toolset for operating and fabricating microfluidic devices
US9365418B2 (en) * 2011-09-02 2016-06-14 The Regents Of The University Of California Universal hardware platform and toolset for operating and fabricating microfluidic devices
US9657286B2 (en) 2012-07-23 2017-05-23 Hitachi High-Technologies Corporation Pre-processing/electrophoresis integrated cartridge, pre-processing integrated capillary electrophoresis device, and pre-processing integrated capillary electrophoresis method
US20170056880A1 (en) * 2015-08-26 2017-03-02 EMULATE, Inc. Fluid connections using guide mechanisms
US20180292428A1 (en) * 2015-10-30 2018-10-11 Konica Minolta, Inc. Liquid delivery method, liquid delivery apparatus and analyzer
US11061046B2 (en) * 2015-10-30 2021-07-13 Konica Minolta, Inc. Liquid delivery method, liquid delivery apparatus and analyzer
US10744502B2 (en) 2016-10-07 2020-08-18 Boehringer Ingelheim Vetmedica Gmbh Analysis device and method for testing a sample
US10953403B2 (en) 2016-10-07 2021-03-23 Boehringer Ingelheim Vetmedica Gmbh Method and analysis system for testing a sample
US20200150142A1 (en) * 2017-06-06 2020-05-14 The Regents Of The University Of California Systems and methods for rapid generation of droplet libraries
US11754579B2 (en) * 2017-06-06 2023-09-12 The Regents Of The University Of California Systems and methods for rapid generation of droplet libraries

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US20070071637A1 (en) 2007-03-29
KR20040088383A (ko) 2004-10-16
CN1912628A (zh) 2007-02-14
CN1912628B (zh) 2011-08-31
KR100755286B1 (ko) 2007-09-04
EP1473084A3 (fr) 2005-02-16
CN1279361C (zh) 2006-10-11
EP1473084A2 (fr) 2004-11-03
US7988913B2 (en) 2011-08-02
EP1473084B1 (fr) 2015-07-29
CN1534297A (zh) 2004-10-06

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