Method and device for measuring multiple physiological properties of cells

• US 9,170,253 B2
• Filed: 03/04/2014
• Issued: 10/27/2015
• Est. Priority Date: 09/10/2003
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Seahorse Bioscience
• Inventors: Teich, Jay S., Neilson, Andy C., Sweeney, Michael R., Uhl, Geoff
• Primary Examiner(s): Shen, Bin
• Application Number:
  US14/196,550
• Publication Number:
  US 20140248650A1
• Time in Patent Office: 602 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0663 Whole sensors
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0851 Bottom walls
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 33/5023 on expression patterns

Abstract

A method of analyzing cells disposed in media within a vessel includes the steps of providing a vessel having an original volume of media about the cells, reducing the original volume of media about at least a portion of the cells to define a reduced volume of media, and analyzing a constituent related to the cells within the reduced volume of media. An apparatus for analyzing cells includes a stage adapted to receive a vessel holding cells and a volume of media, a plunger adapted to receive a barrier to create a reduced volume of media within the vessel including at least a portion of the cells, the barrier adapted for insertion into the vessel by relative movement of the stage and the plunger, and a sensor in sensing communication with the reduced volume of media, wherein the sensor is configured to analyze a constituent disposed within the reduced volume.

Claims

1. A method of determining a cell constituent extracted from or secreted into a volume of media disposed in a well by a sample comprising cells, the method comprising the steps of:

(a) placing in an apparatus a multiwell plate comprising a plurality of said wells, at least one well containing said sample in an original volume of media, the apparatus comprising;

a plurality of moveable barriers that reduce a volume of the media in contact with the samples in the wells; and

sensors disposed on respective surfaces of said barriers in sensing contact with media in said wells,

each barrier and respective sensor disposed on the surface thereof being configured for insertion into a respective well of the multiwell plate;

(b) moving the barriers and sensors into the wells to reduce the volume of the media about the cells in at least one well, defining a reduced-volume sample chamber in contact with the sample;

(c) analyzing with the respective sensor the cell constituent in the media in the reduced-volume sample chamber; and

(d) thereafter, moving the barriers to increase the reduced volume of media about the cells to substantially the original volume.

2. The method of claim 1, wherein analyzing the cell constituent in the media in the reduced-volume sample chamber comprises determining a first concentration of the constituent, and determining a second concentration of the constituent at a predetermined time interval from determining the first concentration.

3. The method of claim 1 wherein the reduced volume is in a range of about 5-50% of the original volume.

4. The method of claim 1 wherein the reduced volume is less than about 5% of the original volume.

5. The method of claim 1 wherein the cells comprise a cell selected from the group consisting of bacteria, fungus, yeast, a prokaryotic cell, a eukaryotic cell, an animal cell, a human cell, and an immortal cell.

6. The method of claim 1 wherein at least a portion of the cells are attached to a surface of the vessel.

7. The method of claim 1 wherein at least a portion of the cells are suspended in the media.

8. The method of claim 1 wherein at least a portion of the cells comprises living tissue.

9. The method of claim 1 wherein the constituent comprises a material selected from the group consisting of a dissolved gas, an ion, a protein, a substrate, a salt, and a mineral.

10. The method of claim 1 wherein the constituent comprises a material extracted from the media by at least a portion of the cells.

11. The method of claim 1 wherein the constituent comprises a material secreted into the media by at least a portion of the cells.

12. The method of claim 1 wherein analyzing the constituent comprises sensing presence of the constituent.

13. The method of claim 1 wherein analyzing the constituent comprises sensing concentration of the constituent.

14. The method of claim 1 wherein analyzing the constituent comprises sensing a first concentration of a first constituent, sensing a second concentration of a second constituent, and determining a relationship between the first concentration and the second concentration.

15. The method of claim 1 wherein analyzing the constituent comprises sensing a rate of change of concentration of the constituent.

16. The method of claim 1 wherein at least one sensor comprises a sensor selected from the group consisting of a fluorescent sensor, a luminescent sensor, an ion-sensitive field-effect transistor (“

ISFET”

) sensor, a surface plasmon resonance sensor, a sensor based on an optical diffraction principle, a sensor based on a principle of Wood'"'"'s anomaly, an acoustic sensor, and a microwave sensor.

17. The method of claim 1 wherein analyzing the constituent comprises determining a parameter selected from the group consisting of cell viability, cell number, cell growth rate, response to at least one of a drug, a toxin, and a chemical, detection of an entity, and internalization.

18. The method of claim 1, further comprising the step of perfusing additional media through the vessel.

19. The method of claim 1, further comprising the step of altering an environment of at least a portion of the cells prior to reducing the original volume of media.

20. The method of claim 1, further comprising the step of altering an environment of at least a portion of the cells after reducing the original volume of media.