Partition-based method of analysis

Patent metadata

Field	Value
Publication number	US-12162008-B2
Application number	US-202117486667-A
Country	US
Kind code	B2
Filing date	Sep 27, 2021
Priority date	Sep 23, 2008
Publication date	Dec 10, 2024
Grant date	Dec 10, 2024

Abstract

Methods of partition-based analysis. In an exemplary method, a device having a port fluidically connected to a chamber may be selected. A sample-containing fluid may be placed into the port. The sample-containing fluid may be moved from the port to the chamber. Partitions of the sample-containing fluid may be formed. A monolayer of the partitions in the chamber may be created. At least a portion of the monolayer may be imaged.

First claim

We claim:

1. A method of analysis, the method comprising:
selecting a device having a port fluidically connected to a chamber;
placing a sample-containing fluid into the port;
moving the sample-containing fluid from the port to the chamber, thus forming sample-containing fluid partitions within the chamber;
forming a monolayer of partitions in the chamber of the sample-containing fluid; and
imaging at least a portion of the monolayer inside the chamber.

2. The method of claim 1 , wherein placing a sample-containing fluid includes placing the sample-containing fluid as a continuous phase into a well of the device.

3. The method of claim 1 , wherein forming partitions includes forming aqueous droplets that are separated from one another by an immiscible carrier liquid.

4. The method of claim 1 , wherein forming partitions includes forming partitions each having a dimension corresponding to a height of the chamber.

5. The method of claim 1 , wherein moving the sample-containing fluid includes creating a pressure differential that drives the sample-containing fluid to the chamber.

6. The method of claim 5 , wherein creating a pressure differential includes applying positive gas pressure or negative gas pressure to a port of the device.

7. The method of claim 1 , further comprising thermally cycling the monolayer of the partitions to promote nucleic acid amplification in a subset of the partitions of the monolayer.

8. The method of claim 1 , wherein imaging at least a portion of the monolayer includes detecting fluorescence from the monolayer.

9. The method of claim 1 , wherein imaging at least a portion of the monolayer includes capturing one or more images, wherein only a subset of the partitions of the monolayer contain an analyte, and wherein the one or more images indicate whether the analyte is present in individual partitions of the monolayer.

10. The method of claim 9 , further comprising determining a number of partitions containing the analyte using the one or more images.

11. The method of claim 9 , wherein the analyte is a nucleic acid target, further comprising amplifying the nucleic acid target within partitions of the monolayer.

12. The method of claim 11 , wherein amplifying the nucleic acid target includes thermally cycling the monolayer to promote nucleic acid amplification in the partitions of the monolayer.

13. The method of claim 11 , wherein the partitions of the monolayer contain an intercalating dye or contain a probe including an oligonucleotide labeled with a fluorophore, and wherein imaging at least a portion of the monolayer includes detecting fluorescence from the intercalating dye or the fluorophore.

14. The method of claim 1 , wherein the chamber lies in a plane and defines an area of the plane, and wherein creating a monolayer of the partitions includes covering a majority of the area with the partitions.

Assignees

• Bio Rad Laboratories Inc

Inventors

• Colston Jr Billy Wayne
• Hindson Benjamin Joseph
• Ness Kevin Dean
• Masquelier Donald Arthur
• Milanovich Fred Paul
• Modlin Douglas N
• Riot Vincent
• Burd Samuel
• Makarewicz Jr Anthony Joseph
• Belgrader Phillip

Classifications

• G01N35/08

  using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• B01L2400/049

  vacuum · CPC title

• B01L2300/0861

  Configuration of multiple channels and/or chambers in a single devices · CPC title

• B01L2300/0829

  Multi-well plates; Microtitration plates · CPC title