Methods for culturing and analyzing cells

• US 9,597,644 B2
• Filed: 03/26/2010
• Issued: 03/21/2017
• Est. Priority Date: 09/05/2003
• Status: Active Grant

• Current Assignee: Stokes Bio Ltd. (Perspecta, Inc.), Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Stokes Bio Ltd. (Perspecta, Inc.)
• Inventors: Davies, Mark, Dalton, Tara
• Primary Examiner(s): Mui, Christine T
• Application Number:
  US12/732,769
• Publication Number:
  US 20100297685A1
• Time in Patent Office: 2,552 Days
• Field of Search: 422/57, 422/50, 422/68.1, 435/309.1, 435/283.1, 435/91.2, 435/91.1, 435/90, 435/85, 435/84, 435/72, 435/41, 436/63
• US Class Current: 1/1
• CPC Class Codes: B01F 25/10 Mixing by creating a vortex flow, e.g. by tangential introduction of flow components
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 2035/00514 Stationary mixing elements
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 35/08 using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G01N 35/1095 for supplying the samples to flow-through analysers for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00

Abstract

Provided herein is a method for analyzing the affect of at least one agent on a cell or cellular component using an analysis system. The method includes wrapping a droplet in an immiscible carrier fluid using a droplet generator wherein the wrapped droplet includes at least one cell or cellular component and at least one agent. The wrapped droplet then flows through the analysis system using a siphoning effect. The droplet is then analyzed to determine effect of the agent on the cell or cellular component.

Claims

1. A method for analyzing a biological sample using an analysis system, the method comprising the steps of:

acquiring the biological sample from a sample source using a sample acquisition device;

transferring the acquired biological sample to a droplet generator, the droplet generator comprising a liquid bridge,

providing a flow of cell culture media to the droplet generator,

mixing the biological sample and cell culture media at the liquid bridge,

segmenting, using the droplet generator, the mixed acquired biological sample and cell culture media into a plurality of individually mixed wrapped droplets, wherein the droplets are wrapped in an immiscible carrier fluid using the droplet generator and comprise at least the biological sample, the cell culture media, and a reagent;

collecting the wrapped sample droplet in a vessel, the vessel configured to contain a carrier fluid for receiving and protecting the wrapped sample droplet;

flowing the wrapped droplet through the analysis system; and

analyzing the wrapped droplet to detect products of a polymerase chain reaction.

2. The method according to claim 1, wherein prior to the analyzing step, the method further comprises dispensing the wrapped droplet to at least one collecting vessel.

3. The method according to claim 1, wherein prior to the analyzing step, the method further comprises lysing the at least one cell to extract a cellular component.

4. The method according to claim 1, wherein prior to the analyzing step, the method further comprises establishing a siphoning effect to dispense the wrapped droplet.

5. The method according to claim 1, wherein the droplet generator is a liquid bridge.

6. A method for analyzing a biological sample using an analysis system, the method comprising the steps of:

acquiring the biological sample from a sample source using a sample acquisition device;

transferring the acquired biological sample to a droplet generator, the droplet generator comprising a liquid bridge;

providing a flow of cell culture media to the droplet generator;

mixing the biological sample and cell culture media at the liquid bridge;

segmenting, using the droplet generator, the mixed acquired biological sample and cell culture media in the droplet generator into a plurality of individual mixed sample droplets wrapped in an immiscible carrier fluid, the sample droplets transferred from the sample acquisition device, wherein the wrapped droplets comprise at least the biological sample, the cell culture media, and a reagent;

flowing the wrapped droplets through the analysis system to detect products of a polymerase chain reaction;

dispensing the wrapped droplets to at least one vessel configured to contain a carrier fluid for receiving and protecting the wrapped sample droplets;

an analysis system for analyzing the wrapped sample droplet and detecting products of a polymerase chain reaction.

7. The method according to claim 6, wherein the droplets comprise a single cell.

8. The method according to claim 6, wherein prior to the dispensing step, the method further comprises lysing the cell in the wrapped droplet.

9. The method according to claim 6, wherein generating comprises:

flowing a sample fluid comprising a plurality of cells to a liquid bridge, wherein the liquid bridge segments the sample fluid to produce the wrapped sample droplets.

10. The method according to claim 6, wherein generating comprises:

dipping an open ended channel into a vessel to produce a wrapped droplet comprising at least one cell.