System and apparatus for sequential processing of analytes

• US 20020051992A1
• Filed: 07/17/2001
• Published: 05/02/2002
• Est. Priority Date: 05/23/1997
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Lynx Therapeutics, Inc. (Illumina Incorporated)
• Inventors: Bridgham, John, Brenner, Sydney, Pallas, Michael C., Corcoran, Kevin P., Golda, George S.
• Granted Patent: US 6,831,994 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0403 specific forces
• CPC Class Codes: B01L 2400/0475 specific mechanical means and fluid pressure
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2521/313 Type II endonucleases, i.e. cutting outside recognition site
• CPC Class Codes: C12Q 2525/161 incorporating target specific and non-target specific sites
• CPC Class Codes: C12Q 2525/191 incorporating an adaptor
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 15/1433 : using image recognition
• CPC Class Codes: G01N 15/1456 : without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 15/1475 : using image analysis for extracting features of the particle
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 15/149 : specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/0015 : in solids
• CPC Class Codes: G01N 2015/149 : Sorting the particles
• CPC Class Codes: G01N 21/253 : for batch operation, i.e. multisample apparatus analytical automats G01N35/00

Abstract

An apparatus and system are provided for simultaneously analyzing a plurality of analytes anchored to microparticles. Microparticles each having a uniform population of a single kind of analyte attached are disposed as a substantially immobilized planar array inside of a flow chamber where steps of an analytical process are carried out by delivering a sequence of processing reagents to the microparticles by a fluidic system under microprocessor control. In response to such process steps, an optical signal is generated at the surface of each microparticle which is characteristic of the interaction between the analyte carried by the microparticle and the delivered processing reagent. The plurality of analytes are simultaneously analyzed by collecting and recording images of the optical signals generated by all the microparticles in the planar array. A key feature of the invention is the correlation of the sequence of optical signals generated by each microparticle in the planar array during the analytical process.

Claims

1. An apparatus for sequential processing of a plurality of analytes, the apparatus comprising:

a flow chamber for disposing a population of microparticles in a planar array, each microparticle of a plurality of microparticles of the population having an analyte anchored thereto;

fluidic means for sequentially delivering processing reagents from one or more reservoirs to the flow chamber so that the analytes anchored to the plurality of microparticles are sequentially exposed to the processing reagents; and

detection means for detecting a sequence of optical signals from each microparticle of the plurality, each optical signal of the sequence being indicative of an interaction between a processing reagent and the analyte anchored thereto.

4. The apparatus of any preceding claim wherein said detection means further comprises an optical train for focusing an image of said optical signals from said planar array of said microparticles onto an electro-optic detector for generating a digital image of said optical signals, and recording means for recording a plurality of digital images of said planar array of said microparticles.

5. The apparatus of claim 4 wherein said detection means further comprises signal tracking means for correlating said optical signals from each of said microparticles in each of said digital images to form for each said microparticle of said plurality a sequence of said optical signals.

6. The apparatus of claim 5 wherein said electo-optical detector is a CCD camera.

7. The apparatus of any preceding claim wherein said analyte is DNA and wherein said optical signal is a fluorescent signal.

2. The apparatus of claim I wherein said flow chamber has an axis and comprises in sequence along the axis an inlet, a planar cavity, a dam, and an outlet such that said processing reagents enter said flow chamber through the inlet and flow in the direction of the axis through the planar cavity and to the outlet.

3. The apparatus of claim I or 2 claim wherein said planar cavity of said flow chamber further has a optically transmissive ceiling and a floor, the optically transmissive ceiling and the floor being parallel to one another and the floor having a plurality of parallel ridges, the parallel ridges being disposed parallel to said axis of said flow chamber, and the parallel ridges being spaced so that said microparticles in said planar cavity form rows between the parallel ridges.