Closed-loop biochemical analyzers

• US 6,391,622 B1
• Filed: 06/27/2000
• Issued: 05/21/2002
• Est. Priority Date: 04/04/1997
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, John Wallace, Kopf-Sill, Anne R., Bousse, Luc J., Knapp, Michael
• Primary Examiner(s): Siew, Jeffrey
• Application Number:
  US09/605,548
• Time in Patent Office: 693 Days
• Field of Search: 435/6, 435/7.1, 435/91.1, 435/91.2, 435/283.1, 435/285.2, 435/287.2, 435/289.1, 536/22.1, 536/23.1, 536/24.3, 536/24.31, 536/24.32, 536/24.33, 204/450, 204/451
• US Class Current: 435/285.2
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/0262 using touch-off at substrate or container
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: 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 B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502753 : characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6869 : Methods for sequencing
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: G01N 2035/1053 : for separating part of the liquid, e.g. filters, extraction phase
• CPC Class Codes: G01N 2035/1058 : for mixing

Abstract

Integrated systems, apparatus, software, and methods for performing biochemical analysis, including DNA sequencing, genomic screening, purification of nucleic acids and other biological components and drug screening are provided. Microfluidic devices, systems and methods for using these devices and systems for performing a wide variety of fluid operations are provided. The devices and systems of are used in performing fluid operations which require a large number of iterative, successive or parallel fluid manipulations, in a microscale, or sealed and readily automated format.

Claims

1. A method of performing a fluidic operation that comprises a plurality of parallel fluid manipulations to provide parallel fluidic analysis of sample materials, the method comprising:

providing an automatable microfluidic device comprising at least a first transverse reagent introduction channel fluidly connected to a source of at least one reagent and a source of at least one sample material, the transverse channel fluidly connected to a plurality of parallel reagent reaction channels, wherein member reagent reaction channels comprise at least one region with a cross sectional dimension of between about 0.1 μ

m and about 500 μ

m;

selecting a first reagent from the source of at least one reagent, transporting the first reagent through the reagent introduction channel and aliquoting a portion of the first reagent into at least one parallel reagent reaction channel;

selecting a first sample material from the source of at least one sample material and aliquoting the first sample material into at least a first of the plurality of parallel reagent reaction channels;

selecting at least one additional sample material, or at least one additional reagent, and aliquoting the additional sample material or additional reagent into at least a second of the plurality of parallel reagent reaction channels;

contacting the first sample material and the first reagent in the first reagent reaction channels, whereupon the first sample material and the first reagent react;

contacting the at least one additional sample material or at least one additional reagent with one or more fluid component selected from the group consisting of the first sample material, the first reagent, the at least one additional reagent, the at least one additional sample material, a second additional reagent, and a second additional sample material;

detecting a first reaction product of the first sample material and the first reagent;

detecting an additional reaction product of the at least one additional sample material or at least one additional reagent and one or more fluid component;

based upon the first or additional reaction product, performing an automated selection of a secondary reagent and a secondary sample material;

introducing the secondary reagent into one of the parallel reaction channels, whereupon the secondary sample material and the secondary reagent reacts; and

,

detecting a secondary reaction product of the secondary sample material and the secondary reagent, thereby providing a fluidic analysis of the first sample material and the secondary sample material.

2. The method of claim 1, wherein the microfluidic device comprises an electrokinetic fluid direction system for moving fluidic components in the device.

3. The method of claim 1, wherein the method comprises parallel analysis of a plurality of sample materials in the parallel channels, in which multiple reagents are mixed in a plurality of the parallel channels with multiple sample materials to form a multiple of products, and, based upon detection of the multiple products, selecting the secondary sample material and secondary reagent.

4. The method of claim 1, wherein the method comprises parallel analysis of a plurality of sample materials in the parallel channels, in which multiple reagents are mixed in a plurality of the parallel channels with multiple sample materials to form a multiple of products, and, based upon detection of the multiple products, selecting multiple secondary sample materials and multiple secondary reagents which are mixed to form multiple reaction products.

5. The method of claim 3, the method comprising parallel analysis of a plurality of sample materials and a plurality of reagents in the parallel channels, in which the plurality of reagents is mixed in a plurality of the parallel channels with a plurality of sample materials to form a plurality of products, and, based upon the plurality of products, selecting a plurality of additional sample materials and additional reagents for subsequent parallel mixing in the parallel channels.

6. The method of claim 1, wherein the microfluidic device includes the first transverse reagent introduction channel and at least a second transverse channel, and a plurality of parallel channels intersecting both of the first and second transverse channels;

and the step of aliquoting the portion of the reagent into at least one parallel reagent reaction channel is performed by;

applying a first voltage across the first transverse reagent introduction channel and the second transverse channel to draw the portion of the reagent into the first transverse reagent introduction channel, whereby the portion of the reagent is present at intersections of the first channel and each of the plurality of parallel channels; and

,

applying a second voltage from the first transverse channel to the second transverse channel, whereby a current in each of the parallel channels is equivalent, and whereby the portion of the reagent at the intersections of the first transverse channel and each of the plurality of parallel channels is moved in to each of the plurality of parallel channels.

7. A method of performing a plurality of separate assays on a single sample, comprising:

providing a microfluidic device having at least a first transverse channel fluidly connected to at least a source of the sample, a plurality of separate parallel channels fluidly connected to the first transverse channel, the member channels comprising at least one reaction region with a cross sectional dimension of between about 0.1 μ

m and about 500 μ

m, and having disposed therein reagents for performing one of a plurality of different diagnostic assays, and a fluid direction system for concurrently directing a portion of the sample into each of the plurality of parallel channels;

transporting a portion of the sample into each of the parallel channels, whereby the sample and the reagents disposed in the at least one region of the channel undergo a reaction;

detecting a result of the reaction of the sample and the reagents for each of the parallel channels;

based upon the result of the reaction, providing each of the plurality of channels with additional reagents for performing one of a plurality of different diagnostic assays; and

repeating the transporting and detecting steps.

8. A method of identifying the presence or absence of a plurality of different predetermined sequence variations at different loci on a target nucleic acid sequence, comprising:

delivering the target nucleic acid sequence to each of a plurality of separate reaction chambers or channels in a microfluidic device, wherein the member chambers or channels comprise at least one reaction region with a cross sectional dimension of between about 0.1 μ

m and about 500 μ

m;

in the reaction region of each of the reaction chambers or channels, amplifying separate regions of the target nucleic acid sequence, each of the separate regions encompassing at least one of the different loci; and

determining whether each of the separate regions of the target nucleic acid sequence contains the sequence variation.

9. The method of claim 8, wherein the step of determining comprises determining a size of the separate region and comparing it to a size of the separate region in the absence of the sequence variation, a difference being indicative of a sequence variation in the region.

10. A method of performing a fluidic operation that requires a plurality of successive fluid manipulations on a sample, comprising:

providing a microfluidic device that comprises at least a first channel fluidly connected to a source of the sample, the first channel being intersected by at least second and third channels, the second and third channels being fluidly connected to a source of first fluid reactant and a source of second fluid reactant, respectively;

transporting a volume of the sample from the source of sample into the first channel;

transporting a volume of the first fluid reactant from the source of first fluid reactant to the first channel to combine with the sample, thereby forming a product; and

transporting a volume of the second fluid reactant from the source of second fluid reactant to the first channel to combine with the product.

11. The method of claim 10, wherein the device comprises an electrokinetic fluid direction system and the transporting steps are performed by electrokinesis.

12. The method of claim 10, wherein the sample is a nucleic acid, the first fluid reactant comprises a sequencing mixture and the second fluid reactant comprises a sequencing stop buffer.

13. The method of claim 10, wherein the first fluid reactant and second fluid reactant are sequencing reagents.

14. The method of claim 10, wherein a product of the combination of the first and second reactants and the sample is detected.

15. The method of claim 10, wherein a product of the combination of the first and second reactants and the sample is detected, wherein results of the detection are used to select a second sample, or to select an additional reactant for subsequent mixing with a component selected from the group consisting of the first reactant, the second reactant, the first sample, the second sample and the additional reactant.

16. An automatable microfluidic device for concurrently performing a plurality of diagnostic assays on one or more samples, comprising:

a plurality of parallel reaction channels, the reaction channels comprising a cross sectional dimension of between about 0.1 μ

m and about 500 μ

m and having disposed therein reagents for performing a diagnostic assay;

a source of the one or more samples;

a sample introduction channel which is fluidly connected to the source of sample and which intersects each of the parallel channels;

a fluid direction system for directing a portion of the sample into each of the plurality of different parallel channels, and

a control system operably coupled to the fluid direction system, wherein the control system comprises an automatable control element for automatically selecting which diagnostic assays are run in the device based upon a detected result from a previous assay performed in the device.

17. The microfluidic device of claim 16, wherein the control system further comprises a control element for selecting a sample from the source of the one or more samples.

18. The microfluidic device of claim 16, wherein the control element selects a first diagnostic assay, detects the results of the first assay and selects a second diagnostic assay based upon the results of the first assay.

19. The microfluidic device of claim 16, wherein the control element selects a first parallel series of diagnostic assays, detects the results of the first parallel series of assays and selects a second series of diagnostic assays based upon the results of the first series of assays.