METHODS FOR USE WITH NANOREACTORS

• US 20100075436A1
• Filed: 05/06/2009
• Published: 03/25/2010
• Est. Priority Date: 05/06/2008
• Status: Abandoned Application

• Current Assignee: Tethys Bioscience, Inc.
• Original Assignee: Tethys Bioscience, Inc.
• Inventors: URDEA, Michael S., TARASOW, Theodore M., MOLER, Edward J.
• Application Number:
  US12/436,585
• Publication Number:
  US 20100075436A1
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• 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 2300/0896 Nanoscaled
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites

Abstract

The invention relates to methods of using nanoreactor technology for sample analysis in microfluidic systems.

Claims

1. A method of washing a nanoreactor containing a particle in a microfluidic system, comprising the steps of:

a) fusing a first nanoreactor containing a particle with a second nanoreactor containing a washing solution to form a combined nanoreactor, wherein the diameter of the second nanoreactor is at least about two fold the diameter of the first nanoreactor; and

wherein a molecule within the first nanoreactor is diluted in the combined nanoreactor;

b) splitting the combined nanoreactor into a plurality of nanoreactors; and

c) separating the nanoreactor containing the particle from the plurality of nanoreactors formed in step b) in a microfluidic system.

2. The method of claim 1, wherein the diameter of the second nanoreactor is at least about five fold of the diameter of the first nanoreactor.

3. The method of claim 1, wherein the diameter of the second nanoreactor is at least about ten fold of the diameter of the first nanoreactor.

4. The method of claim 1, wherein the particle comprises a reporter

5. The method of claim 4, wherein the reporter is a dye coded bead or a nano-bar code.

6. The method of claim 1, wherein the first nanoreactor has a cross-sectional dimension of less than about 100 microns.

7. The method of claim 1, wherein the first nanoreactor has a cross-sectional dimension of less than about 30 microns.

8. The method of claim 1, wherein the first nanoreactor has a cross-sectional dimension of less than about 10 microns.

9. The method of claim 1, wherein the first nanoreactor has a cross-sectional dimension of less than about 3 microns.

10. The method of claim 1, wherein the method is used in a washing step of a heterogeneous assay.

11. The method of claim 1, wherein the plurality of nanoreactors which do not contain the particle are returned to a starting pool for further analysis.

12. A method for tracking a sample in a nanoreactor comprising the steps of a) fusing a sample nanoreactor comprising a sample and a reporter with a reagent nanoreactor comprising a particle and a reagent, wherein the reporter comprises a first reactive group, and a second reactive group and a reagent are associated with the particle;

wherein the first reactive group reacts with the second reactive group so that the reporter is linked to the particle; and

b) tracking the sample nanoreactor that has reacted with the reagent nanoreactor by tracking the nanoreactor containing the reporter.

13. The method of claim 12, wherein the reporter is covalently linked to the particle in step a).

14. The method of claim 12, wherein the reporter is not covalently linked to the particle in the step a).

15. The method of claim 12, wherein the reporter is selected from the group consisting of a dye, a fluorescent agent, an ultraviolet agent, a chemiluminescent agent, a chromophore, a radio-label, a mass spectrometry tag molecule, and a resonance raman tag molecule.

16. A method of measuring concentration of an analyte in a sample, said method comprising:

a) compartmentalizing a sample into a plurality of nanoreactors, wherein at least about 80% of the nanoreactors contain no more than a single analyte molecule; and

b) detecting the nanoreactor containing at least an analyte molecule;

wherein the number of nanoreactors containing analyte molecules indicates the concentration of the analyte in the sample.

17. The method of claim 16, wherein at least about 90% of the nanoreactors contain no more than a single analyte molecule.

18. The method of claim 16, wherein at least about 95% of the nanoreactors contain no more than a single analyte molecule.

19. The method of claim 16, wherein greater than 95% of the nanoreactors contain no more than a single analyte molecule.

20. The method of claim 16, wherein the analyte containing nanoreactors are detected by labeling the analyte with a reporter.

21. The method of claim 16, wherein the concentration of the analyte in the sample is about 5 aM to about 500 fM.

22. The method of claim 16, wherein the analyte is selected from the group consisting of a protein, a peptide, an oligonucleotide, a metabolite, a carbohydrate, a lipid, a ligand, a receptor, and a small molecule.

23. The method of claim 16, wherein the sample is a clinical sample selected from the group consisting of blood, plasma, serum, saliva, urine, and spinal fluid.

24. The method of claim 16, wherein the nanoreactors have a cross-sectional dimension of less than about 100 microns.

25. The method of claim 16, wherein the nanoreactors have a cross-sectional dimension of less than about 30 microns.

26. The method of claim 16, wherein the nanoreactors have a cross-sectional dimension of less than about 10 microns.