Device and methods for molecular analysis

Patent metadata

Field	Value
Publication number	US-9605298-B2
Application number	US-201013389259-A
Country	US
Kind code	B2
Filing date	Aug 6, 2010
Priority date	Aug 6, 2009
Publication date	Mar 28, 2017
Grant date	Mar 28, 2017

Abstract

Systems and methods are provided for high speed sorting of objects in a continuous body of fluid. The object can be analyzed within one or more interrogation volumes that allow for simultaneous or time-correlated measurement of the object's properties. A processor can interpret the properties of the object and then measured and then direct the object to one of a plurality of downstream flow paths. In some embodiments, the sorting of the object is based on two or more properties of the object. The sorting process can be repeated to create a network of sorting events.

First claim

What is claimed is:

1. A system for sorting chromatin comprising:
(a) a channel that is fluidically connected to a plurality of downstream flow paths, said channel being adapted to hold said chromatin in a continuous liquid body in said channel;
(b) one or more light sources configured to illuminate the channel to create one or more interrogation volumes, wherein the one or more interrogation volumes are bounded by a submicrometer channel region of said channel, and wherein the one or more interrogation volumes are each less than 0.5 femtoliters;
(c) a detection module configured to detect at least two types of signals indicative of two distinct properties of the chromatin in the one or more interrogation volumes;
(d) a sorting module configured to direct the chromatin to one of the plurality of downstream flow paths based on the two distinct properties indicated by said two types of signals; and
(e) one or more electrodes in electrical connection by fluid with the channel and downstream flow paths to direct the chromatin.

2. The system of claim 1 , further comprising:
(a) one or more secondary light sources configured to illuminate one of the plurality of downstream flow paths to create one or more downstream interrogation volumes; and
(b) a secondary detection module configured to detect at least one type of signals in the one or more downstream interrogation volumes.

3. The system of claim 2 , further comprising a secondary sorting module configured to direct the chromatin to one of a plurality of secondary downstream flow paths based on said two types of signals in the one or more downstream interrogation volumes, wherein the plurality of secondary downstream flow paths are different from the plurality of downstream flow paths of (a).

4. A method for sorting an object in a channel, comprising:
(a) flowing the object through said channel, wherein said object is labeled with a plurality of labels;
(b) illuminating the channel to create one or more interrogation volumes, each of which is confined by walls of a submicrometer channel region of said channel and a beam of light, and wherein each of said one or more interrogation volumes has a volume of less than 0.5 femtoliters; and
(c) detecting a first label and a second label of said plurality in the same or distinct interrogation volumes of said one or more interrogation volumes to generate time-correlated resolution of said first and second label,
wherein the first label is indicative of a first property of the object and the second label is indicative of a second property of the object; and

(d) directing the object to one of a plurality of downstream flow paths that are fluidically connected to said channel, wherein the object is directed based on the first property and the second property, and wherein the object is directed by electrodes in electrical connection by fluid with the channel and downstream flow paths.

5. The method of claim 4 , further comprising:
(a) illuminating a downstream pathway to create one or more downstream interrogation volumes; and
(b) detecting at least one label of said plurality in the one or more downstream interrogation volumes.

6. The method of claim 5 , further comprising directing the object to one of a plurality of secondary downstream flow paths based on said detecting at least one label in the one or more downstream interrogation volumes, wherein the plurality of secondary downstream flow paths are different from the plurality of downstream flow paths of (d).

7. The method of claim 4 , wherein the object is a chromatin.

8. The system of claim 1 , wherein the one or more interrogation volumes are each less than 0.2 femtoliters.

9. The system of claim 1 , wherein the sorting module has a switching time that is less than 100 microseconds.

10. The system of claim 1 , wherein the one or more electrodes are capable of imparting an electrokinetic force on the object.

11. The system of claim 1 , wherein the channel and each of the plurality of downstream flow paths has a width less than 1 μm.

12. The system of claim 1 , wherein the channel and the plurality of downstream flow paths each has a width less than 1 μm and a height less than about 1 μm.

13. The method of claim 4 , wherein the one or more interrogation volumes are each less than 0.2 femtoliters.

14. The method of claim 4 , wherein the object is directed by a switching module having a switching time that is less than 100 microseconds.

15. The method of claim 4 , wherein the object is directed by electrokinetic force using the electrodes.

16. The method of claim 4 , wherein the channel and each of the plurality of downstream flow paths has a width less than 1 μm.

17. The method of claim 4 , wherein the channel and the plurality of downstream flow paths each has a width less than 1 μm and a height less than 1 μm.

18. The method of claim 4 , wherein the object is a chromatin, and the first label is complexed with an epigenetic marker on the chromatin and the second label is complexed with a protein or nucleotide of the chromatin.

19. The method of claim 4 , wherein the object is directed less than 0.001 seconds after the first label and second label are detected.

20. A method for sorting an object in a channel, comprising:
(a) flowing the object through said channel, wherein said object is labeled with a plurality of labels;
(b) illuminating the channel to create one or more interrogation volumes, wherein each of said one or more interrogation volumes is confined by walls of a submicrometer channel region of said channel and a beam of light, and each of said one or more interrogation volumes has a volume of less than 0.5 femtoliters; and
(c) detecting a first label and a second label of said plurality of labels in the same or distinct interrogation volumes of said one or more interrogation volumes to generate time-correlated resolution of said first label and second label,
wherein the first label is indicative of a first property of the object and the second label is indicative of a second property of the object; and

(d) directing the object to one of a plurality of downstream flow paths that are fluidically connected to said channel, wherein the object is directed based on the first property and the second property, and further wherein the object is directed less than 0.001 seconds after the first label and second label are detected.

21. The method of claim 20 , wherein the channel and the plurality of downstream flow paths each have a width less than 1 μm and a height less than 1 μm.

22. The system of claim 1 , wherein a branch point of the plurality of downstream flow paths is positioned at or immediately downstream of the detection module.

23. The system of claim 1 , wherein the system is configured such that the sorting module directs the chromatin to one of the plurality of downstream flow paths in less than 0.001 seconds after the at least two types of signals are detected by the detection module.

Assignees

• Craighead Harold G
• Cipriany Benjamin R
• Levy Stephen
• Soloway Paul
• Univ Cornell

Inventors

• Craighead Harold G
• Cipriany Benjamin R
• Levy Stephen
• Soloway Paul

Classifications

• B01L3/502761 Primary

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L2200/0663

  Stretching or orienting elongated molecules or particles · CPC title

• C12Q2561/113

  Real time assay · CPC title

• C12Q2537/164

  Methylation detection other then bisulfite or methylation sensitive restriction endonucleases · CPC title

• B01L2300/0654

  Lenses; Optical fibres · CPC title