Methods and Devices For Detection and Identification of Encoded Beads and Biological Molecules

• US 20090203148A1
• Filed: 01/19/2007
• Published: 08/13/2009
• Est. Priority Date: 01/19/2006
• Status: Abandoned Application

• Current Assignee: The Research Foundation for The State University of New York (State University of New York)
• Original Assignee: The Research Foundation for The State University of New York (State University of New York)
• Inventors: Gorbovitski, Boris, Gorfinkel, Vera, Gorbovitski, Michael
• Application Number:
  US12/087,812
• Publication Number:
  US 20090203148A1
• Time in Patent Office: Days
• US Class Current: 436/94
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The invention relates to methods and devices used in the sequencing, separation, detection, and identification of objects and biological molecules. In preferred embodiments, the invention relates to a DNA sequencing system based on cyclic sequencing by synthesis which is performed on beads in three-dimensional vessels and detected using monolithic multicapillary arrays. In other embodiments, the invention relates to a bead comprising two or more luminescent labels coupled to a nucleic acid sequence. In further embodiments, said luminescent labels are quantum dots.

Claims

10. A detection system comprising:

a) a first bead comprising a first luminescent label and a second luminescent label;

b) a second bead comprising a third luminescent label and a fourth luminescent label;

c) a first transparent channel capable of accepting said first bead and a second transparent channel capable of accepting said second bead; and

d) an instrument for detecting electromagnetic radiation from luminescent labels.

11. The system of claim 10, wherein said first and second luminescent labels are fluorescent quantum dots.

12. The system of claim 10, wherein said first luminescent label and said third luminescent label are the same label wherein said third luminescent label is at lower concentration per bead inside said second bead than is the concentration per bead of said first luminescent label inside said first bead.

13. The system of claim 10, wherein a common wall separates said first and second transparent channels

14. The system of claim 10, wherein said first and second transparent channels comprise a square cross section.

15. The system of claim 10, wherein said instrument for detecting electromagnetic radiation comprises a charge-coupled device.

16. The system of claim 10, further comprising a source of electromagnetic radiation.

17. The system of claim 16, wherein said source of electromagnetic radiation is a laser.

18. A method of generating luminescent encoded beads comprising:

a) providing;

i) a plurality of first luminescent particles,

ii) a plurality of second luminescent particles, and

iii) a plurality of porous structures;

iv) a first plurality of wells wherein

said first luminescent particles are in said wells and have different concentrations in at least two of the wells;

v) a second plurality of wells

wherein said second luminescent particles are in said wells and have different concentrations in at least two of the wells;

b) distributing a portion of said plurality of porous structures to said first plurality of wells under conditions such that said first luminescent particles are absorbed by said porous structures;

c) extracting said plurality of porous structures with said first luminescent particles from said first plurality of wells;

d) mixing said extracted plurality of porous structures with said first luminescent particles together to form a plurality of porous structures wherein at least two of said porous structures have different concentrations of said first luminescent particles;

e) distributing said plurality of porous structures wherein at least two of said porous structures have different concentrations of said first luminescent particles to said second plurality of wells under conditions such that said second luminescent particles are absorbed by said porous structures;

f) extracting said plurality of porous structures with said first luminescent particles and said second luminescent particles from said second plurality of wells

g) mixing said plurality of porous structures with said first luminescent particles and said second luminescent particles that are extracted from said second plurality of wells together to form a plurality of porous structures wherein at least two of said porous structures have different concentrations of said first luminescent particles and have different concentrations of said second luminescent particles.

19. The method of claim 18, wherein said first luminescent particle is a quantum dot.

20. The method of claim 18, wherein said second luminescent particle is a quantum dot wherein said first and second quantum dot have a different size.

21. The method of claim 18, wherein said porous structures are mesoporous silica beads.

22. The method of claim 18, wherein said porous structures are mesoporous polystyrene beads.

23. The method of claim 18, wherein said conditions for distributing a portion of said plurality of porous structures to said first plurality of wells does not saturate the porous structures with said first plurality of particles.

24. The method of claim 18, further providing a plurality of third luminescent particles, wherein said second and third luminescent particles are in said wells and have different concentrations in at least two of the wells and further mixing said plurality of porous structures with said first luminescent particles said second luminescent particles and said third luminescent particles that are extracted from said second plurality of wells together to form a plurality of porous structures wherein at least three of said porous structures have different combinations of concentrations of said first, said second and said third luminescent particles.

25. A method of determining the authenticity of an object comprising

a) providing

i) an object comprising plurality luminescent encoded beads,

wherein said encoded beads comprise two or more luminescent markers configured to provide a luminescent signature,

ii) electromagnetic radiation, and

iii) an instrument for detecting electromagnetic radiation;

b) placing said object in said electromagnetic radiation under conditions such that said quantum dots luminesce, and

c) detecting said luminescent signature with said instrument; and

d) correlating the luminescent signature with the authenticity of said object.

26. The method of claim 25, wherein said object is selected from the group consisting of a personal identification card, cash, liquid, solid, and fabric.

27. The method of claim 25, wherein said electromagnetic radiation is ultraviolet light.

28. The method of claim 25, wherein said luminescent markers are quantum dots.

29. A method of determining a DNA sequence comprising:

a) providing,

i) a plurality of linked beads wherein said beads comprise

A) luminescent electromagnetic codes,

B) a plurality of identical single stranded nucleic acid molecules

wherein said plurality of nucleic acids molecules are configured such that an individual bead having an individual code carries nucleic acids molecules with a unique sequence;

ii) a plurality of wells at least four of said wells contain solutions with four different fluorescently labeled nucleotides;

b) detecting said luminescent electromagnetic codes on said plurality of beads and recording said codes;

c) placing said linked beads into one of said wells containing one of said free nucleotides and incubating said beads under conditions such that said free nucleotides are attached to said nucleic acids molecules to form a complementary strand;

d) detecting said linked beads and determining said codes of said beads for which the incorporation of said free nucleotides occurs;

e) recording in a computer file said codes of said beads for which the incorporation of said free nucleotides occurs;

f) removing fluorescence label from said incorporated free nucleotides;

g) repeatedly repeating steps c), d), e), and f) for all said free nucleotides;

h) analyzing said recorded data on said incorporated nucleotides for each individual bead and determining sequence of said nucleic acid molecules linked to said individual bead.

30. A method of moving a bead through a channel comprising:

a) providing;

i) bead comprising a first luminescent label and a second luminescent label,

ii) a channel,

iii) a solution inside said channel wherein said beads are inside said solution,

iv) pair of electrodes; and

b) applying a potential between said pair of electrodes under conditions such that said bead moves in said channel toward one electrode of said electrode pair.

31. The method of claim 31, wherein said bead is a polystyrene bead.

32-1. The method of claim 32, wherein said bead is charged.