Fluid processing device with captured reagent beads

• US 20060228734A1
• Filed: 03/17/2006
• Published: 10/12/2006
• Est. Priority Date: 03/18/2005
• Status: Abandoned Application

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Ulmanella, Umberto, Vann, Charles S.
• Application Number:
  US11/378,559
• Publication Number:
  US 20060228734A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502761 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 investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 33/54386 : Analytical elements

Abstract

A fluid processing device, method and system are provided. The fluid processing device can comprise: a substrate; a plurality of reaction regions disposed in or on the substrate; at least one channel interconnecting the plurality of reaction regions, the at least one channel having a cross-sectional area that includes a maximum dimension; and a plurality of reagent-releasing beads. Each reagent-releasing bead can be positioned in a respective one of the reaction regions. Each bead can comprise one or more reaction components for an assay. Each of the reagent-releasing beads can have a minimum dimension that is greater than the maximum dimension of the channel cross-section.

Claims

1. A fluid processing device, comprising:

a substrate;

a plurality of reaction regions disposed in or on the substrate;

at least one channel interconnecting the plurality of reaction regions, the at least one channel having a cross-sectional area that includes a maximum dimension; and

a plurality of reagent-releasing beads, each reagent-releasing bead being positioned in a respective one of the reaction regions and comprising one or more reaction components for an assay, wherein each of the reagent-releasing beads has a minimum dimension that is greater than the maximum dimension of the channel cross-section.

2. The fluid processing device of claim 1, further comprising a loading port in fluid communication with the at least one channel.

3. The fluid processing device of claim 2, wherein the volume of the loading port is greater than the total volume of all of the plurality of reaction regions and the plurality of channels, combined.

4. The fluid processing device of claim 2, wherein the at least one channel has a first end and a second end, the loading port is in fluid communication with the first end, and the device further comprises a suction port in fluid communication with the second end.

5. The fluid processing device of claim 4, further comprising a syringe adapted to create suction at the suction port and forming an airtight seal with the suction port.

6. The fluid processing device of claim 4, wherein the at least one channel comprises a plurality of channels, each of the plurality of channels is in fluid communication with a respective plurality of the plurality of reaction regions, and each of the plurality of channels has a first end in fluid communication with the loading port and a second end in fluid communication with the suction port.

7. The fluid processing device of claim 1, wherein each reagent-releasing bead comprises a material that is solid at 25°

C. and dissolves in water at a temperature greater than about 50°

C.

8. The fluid processing device of claim 1, wherein the substrate comprises a top surface and the device further comprises a cover layer that contacts the top surface and encloses the plurality of reaction regions and the at least one channel.

9. The fluid processing device of claim 8, wherein the cover layer comprises a material that is non-porous, gas-permeable, and liquid-impermeable at pressures of 75 pounds per square inch or less.

10. The fluid processing device of claim 1, wherein the substrate comprises a bottom surface and the device further comprises a heat conductive layer having a thermal conductivity of 0.25 Kelvin Watts per meter, that contacts the bottom surface.

11. The fluid processing device of claim 1, wherein the substrate comprises a bottom surface and the device further comprises a cover layer that contacts the bottom surface and encloses at least one of the plurality of reaction regions or the at least one channel.

12. The fluid processing device of claim 2, wherein the at least one channel comprises a plurality of channels, each of the plurality of channels is in fluid communication with a respective plurality of the plurality of reaction regions, and each of the plurality of channels has a first end in fluid communication with the loading port and a second end in fluid communication with a vent.

13. The fluid processing device of claim 1, wherein each of the reagent-releasing beads comprises one or more components for real-time fluorescence-based measurements of nucleic acid amplification products held in at least one of the plurality of reaction regions.

14. The fluid processing device of claim 1, wherein one of the plurality of reagent-releasing beads comprises first and second oligonucleotide primers having sequences effective to hybridize to opposite end regions of complementary strands of a selected polynucleotide analyte segment, for amplifying the segment by primer-initiated polymerase chain reaction, and a fluorescer-quencher oligonucleotide capable of hybridizing to a analyte segment in a region downstream of one of the primers, for producing a detectable fluorescent signal when an analyte is present in an sample.

15. The fluid processing device of claim 1, wherein the at least one channel comprises a plurality of segments for interconnecting the plurality of reaction regions.

16. The fluid processing device of claim 15, further comprising a vent in fluid communication with one end of the at least one channel and a loading port in fluid communication with a distal end of the at least one channel.

17. The fluid processing device of claim 15, further comprising a pressure source adapted to interface with the loading port and capable of injecting a first fluid through the at least one channel and the plurality of reaction regions, while replacing a second fluid therein by venting the second fluid from the vent.

18. The fluid processing device of claim 17, wherein the first fluid comprises a liquid and the second fluid comprises a gas.

19. A system comprising:

a thermal cycler; and

a fluid processing device of claim 1 disposed in the thermal cycler.

20. A system comprising:

a fluid processing device of claim 1 disposed in the thermal cycler; and

a fluorescence detection system adapted to perform real-time polymerase chain reaction detection for one of the plurality of reaction wells.

21. A fluid processing device, comprising:

a substrate; and

a pathway disposed in or on the substrate comprising;

a loading port,

a vent,

a first fluid retainment region comprising a reagent-releasing bead in fluid communication with the loading port,

a second fluid retainment region comprising a reagent-releasing bead in fluid communication with the vent, and

a first channel in fluid communication with the first fluid retainment region and the second fluid retainment region.

22. A method comprising:

loading a fluid processing device with a fluid, wherein the fluid processing device comprises a plurality of reaction regions disposed on or in a substrate, interconnected by at least one channel, and each reaction region comprises a reagent-releasing bead comprising a reagent.

23. The method of claim 22, further comprising releasing reagent from each reagent-releasing bead.

24. The method of claim 22, further comprising carrying out a reaction process in each of the reaction regions.

25. The method of claim 22, wherein the at least one channel comprises a plurality of segments interconnecting the plurality of reaction regions, each segment having a length long enough to prevent interaction of the reagent in one reaction region of the plurality of reaction regions with the reagent released from another reaction region of the plurality of reaction regions.

26. The method of claim 22, further thermal-cycling the fluid processing device.