Flat-field imaging system and methods of use

Patent metadata

Field	Value
Publication number	US-11965809-B2
Application number	US-202117525477-A
Country	US
Kind code	B2
Filing date	Nov 12, 2021
Priority date	Jun 17, 2011
Publication date	Apr 23, 2024
Grant date	Apr 23, 2024

Abstract

A method of aligning a plurality of targets is provided. The method includes generating a plurality of targets. A third phase includes the plurality of targets. The method further includes combining a first phase, a second phase, and the third phase in a volume. The first phase, the second phase, and the third phase are substantially immiscible, and the third phase is in fluid communication with the first phase and the second phase, and the first phase, the second phase, and the third phase are operable to be in a configuration of the third phase between the first phase and the second phase in the volume.

First claim

What is claimed is:

1. A method for analyzing targets comprising biological sample, the method comprising:
forming a volume, wherein the volume comprises a third phase in fluid communication with a first phase and a second phase and is within a field of view of an optical sensor, and an interface between the first phase and the second phase,
wherein the third phase is disposed along the interface,
wherein the third phase includes a plurality of targets comprising biological sample,
wherein the plurality of targets are disposed in a substantially non-overlapping single layer within the volume;
wherein the first phase and the second phase are liquids,
wherein the first phase has a first density, the second phase has a second density, and the third phase as a third density, wherein the first density, the second density, and the third density are different densities, and
wherein the first phase, the second phase, and the third phase are substantially immiscible,
amplifying the biological sample; and
imaging, using the optical sensor, the plurality of targets, to determine amplification of the biological sample.

2. The method of claim 1 , further comprising detecting a number of targets with a positive amplification and a number of targets with a negative amplification within the plurality of targets.

3. The method of claim 2 , further comprising quantifying an amount of biological sample within the plurality of targets.

4. The method of claim 1 , wherein the first density is heavier than the third density and the third density is heavier than the second density.

5. The method of claim 1 , wherein the plurality of targets have substantially the same density as the third density.

6. The method of claim 1 , wherein the first phase comprises a fluorinated fluid (HFE), and the second phase comprises a mineral oil.

7. The method of claim 1 , wherein the plurality of targets comprises targets of a plurality of sizes.

8. The method of claim 1 , wherein the plurality of targets comprise droplets.

9. The method of claim 8 , further comprising agitating the volume including the first phase, second phase, and third phase to generate the plurality of droplets.

10. A system for analyzing targets comprising biological sample, the system comprising:
a volume, wherein the volume comprises: a third phase in fluid communication with a first phase and a second phase, and an interface between the first phase and the second phase,
wherein the first phase and the second phase are liquids,
wherein the first phase has a first density, the second phase has a second density, and the third phase as a third density, wherein the first density, the second density, and the third density are different densities,
wherein the first phase, the second phase, and the third phase are substantially immiscible,
wherein the third phase includes a plurality of targets comprising biological sample,
wherein the third phase is disposed along the interface in a substantially non-overlapping layer within the volume, and
wherein the plurality of targets are disposed in a substantially non-overlapping single layer within the volume;

a thermo cycler for amplifying the biological sample; and
an optical sensor having a field of view including the third phase for imaging the plurality of targets in the volume.

11. The system of claim 10 , wherein the optical sensor is further configured to detect a number of targets with a positive amplification of the biological sample and a number of targets with a negative amplification of the biological sample within the plurality of targets.

12. The system of claim 11 , further comprising a processor configured to quantify an amount of biological sample within the plurality of targets.

13. The system of claim 10 , wherein the first density is heavier than the third density and the third density is heavier than the second density.

14. The system of claim 10 , wherein the plurality of targets have substantially the same density as the third density.

15. The system of claim 10 , wherein the plurality of targets comprises targets of a plurality of sizes.

16. The system of claim 10 , wherein the plurality of targets comprise droplets.

17. The system of claim 10 , wherein at least some of the plurality of targets are porous beads.

18. The system of claim 10 , wherein the plurality of targets are magnetic beads.

19. The system of claim 10 , wherein the third phase is disposed along the interface in a planar layer within the volume.

Assignees

• Life Technologies Corp

Inventors

• Andersen Mark
• Pallas Michael
• Wang Haopeng

Classifications

• G01N15/04 Primary

  Investigating sedimentation of particle suspensions · CPC title

• C12Q1/6816 Primary

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C12Q2563/107

  fluorescence · CPC title

• C12Q2563/159

  Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components · CPC title