Method and system for detecting a target within a population of molecules

Patent metadata

Field	Value
Publication number	US-9575068-B2
Application number	US-201313909109-A
Country	US
Kind code	B2
Filing date	Jun 4, 2013
Priority date	Oct 27, 2006
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

A method of detecting a target within a population of molecules comprising: contacting a plurality of labeled probe molecules with the population of molecules potentially containing a target of the probe molecules; acquiring a probe specific signal emitted by said labeled probe molecules that bound to said target together with a background signal; preferentially modulating said probe specific signal by at least one of modulating said acquisition and modulating an emission of said probe specific signal; and detecting said probe specific signal over said background signal using said preferential modulation.

First claim

What is claimed is:

1. A method of detecting at least one target molecule within a population of molecules, the method comprising:
contacting a plurality of labeled probe molecules, each selectively binds to at least one target molecule, with the population of molecules potentially containing a target of said at least one target molecule;
directing an excitation beam in a first direction through the plurality of probe molecules, thereby producing a background signal and potentially a probe specific signal emitted if at least one of said plurality of labeled probe molecules is bound to at least a portion of said target;
detecting said probe specific signal over said background signal by performing a preferential modulation to at least one of said acquisition and an emission of said probe specific signal, wherein said preferential modulation includes moving the at least one of said plurality of labeled probe molecules bound to at least a portion of said target in a second direction, in and out of the excitation beam, wherein said detecting according to said probe specific signal includes detecting relative movement between the excitation beam and the target molecule, wherein said relative movement results in the labeled probe molecules bound to at least the portion of the target being positioned in and out of the excitation beam, and;
detecting at least one of the presence, the absence and the amount of said at least one target molecule in said target according to said probe specific signal.

2. A method according to claim 1 , wherein said detecting according to said probe specific signal includes detecting an expected Bragg diffraction/scattering of the excitation beam by the target molecule.

3. A method according to claim 1 , wherein the modulation is achieved by spatial arrangement of said at least one target in a pattern.

4. A method according to claim 3 , wherein said pattern is arranged by one of a standing acoustic wave and centrifugation.

5. The method of claim 1 , wherein said preferential modulation comprises temporal modulation.

6. The method of claim 1 , wherein said preferential modulation comprises spatial modulation.

7. The method of claim 1 , wherein said preferential modulation comprises multiple modulation cycles.

8. The method of claim 1 , wherein said preferential modulation comprises a modification of a response of said at least one labeled probe molecule to excitation energy applied thereon.

9. The method of claim 1 , wherein said detecting comprises applying an alternating field for establishing a periodic motion of said at least one bound probe molecule.

10. The method of claim 1 , wherein said plurality of probe molecules is attached to a magnetic particle.

11. The method of claim 1 , wherein the probe specific signal is a fluorescent signal.

12. The method of claim 1 , wherein the detecting said probe specific signal over said background signal is accomplished within one minute of the contacting a labeled probe molecule with the population of molecules.

13. The method of claim 1 , wherein the detecting comprises at least one detection type selected from a group consisting of binary detection, amplitude detection and synchronous detection.

14. The method of claim 1 , wherein said detecting is performed if said preferential modulation produces a probe specific signal being ten times greater in amplitude than the background signal.

15. The method of claim 1 , wherein the preferential modulation comprises increasing a local concentration of said plurality of labeled probe molecules during said detecting.

16. The method of claim 1 , wherein said moving the excitation beam includes moving the excitation beam while the target remains stationary.

17. The method of claim 16 , wherein said moving the excitation beam relative to the target molecule is additionally affected by the target molecule being moved by an acoustic wave.

18. The method of claim 1 , wherein said performing a preferential modulation increases the number of labeled probe molecules being in the excitation beam at one time when the excitation beam is moved.

19. The method of claim 18 , wherein said increase in the number of labeled probe molecules results in an increase in the probe specific signal.

20. The method of claim 1 , wherein said moving the excitation beam does not affect background signal.

21. The method of claim 1 , wherein said detecting includes detecting reflections of the excitation beam from aggregated beads carried on the probe molecules relative to which the excitation beam is moved.

22. A system for analysis of a population of molecules, comprising:
a vessel adapted to contain a plurality of labeled probe molecules, each selectively binds to at least one target molecule, in contact with a population of molecules potentially containing a target comprising said at least one target molecule so that at least one of said plurality of labeled probe molecules being bound to at least a portion of said target;
a fluorescent excitation source adapted to direct an excitation beam in a first direction through the vessel, the beam configured to cause at least some of said at least one labeled probe molecule to emit a probe specific signal;
a detector adapted to detect said probe specific signal and produce a detection output accordingly;
a signal modulator configured to perform a preferential modulation to at least one of the probe specific signal and said detection output, wherein said preferential modulation includes moving the at least one of said plurality of labeled probe molecules bound to at least a portion of said target in a second direction, in and out of the excitation beam; and
an analysis module adapted to analyze the detection output in consideration of said preferential modulation, wherein the detection includes detecting by analyzing a signal provided by the excitation beam when there is relative movement between the excitation beam and the at least one bound probe molecule, wherein said relative movement results in the probe molecules bound to the target being positioned in and out of the excitation beam.

23. The system of claim 22 , comprising a modulation indication source adapted to provide a modulation indication to the signal modulator and to the detector.

24. The system of claim 22 , wherein the signal modulator is adapted to:
monitor an output of the signal modulator; and
provide a modulation indication to said detector responsive to the output of the signal modulator.

25. The system of claim 22 , wherein said detection output includes at least one of the presence, the absence and the amount of said at least one target molecule in said target according to said probe specific signal.

26. The system of claim 25 , wherein said detection output includes an expected Bragg diffraction/scattering of the excitation beam by the target molecule.

27. The system of claim 22 , wherein said signal modulator is configured to achieve modulation by spatial arrangement of said at least one target in a pattern.

28. The system of claim 27 , wherein said pattern is arranged by one of a standing acoustic wave and centrifugation.

29. The system of claim 22 , wherein said preferential modulation comprises temporal modulation.

30. The system of claim 22 , wherein said preferential modulation comprises spatial modulation.

Assignees

• Ramot At Tel-Aviv Univ Ltd

Inventors

• Arie Ady
• Danielli Amos

Classifications

• G01N33/582 Primary

  with fluorescent label · CPC title

• G01N21/6428 Primary

  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 title

• G01N2021/6439

  with indicators, stains, dyes, tags, labels, marks · CPC title

• G01N21/64

  Fluorescence; Phosphorescence · CPC title

• G01N33/54333 Primary

  Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction · CPC title