Cell analysis apparatus and method

• US 9,170,255 B2
• Filed: 01/08/2014
• Issued: 10/27/2015
• Est. Priority Date: 07/13/2006
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Seahorse Bioscience
• Inventors: Teich, Jay S., Neilson, Andy C., Young, Stephen G., Orrell, Jim
• Primary Examiner(s): Shen, Bin
• Application Number:
  US14/150,423
• Publication Number:
  US 20140186876A1
• Time in Patent Office: 657 Days
• Field of Search: 435/287.1
• US Class Current: 1/1
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/047 Additional chamber, reservoir
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/42 Integrated assemblies, e.g. cassettes or cartridges
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/6434 Optrodes
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 2035/0091 GUI [graphical user interfaces]
• CPC Class Codes: G01N 2035/0094 optimisation; experiment design
• CPC Class Codes: G01N 21/03 Cuvette constructions
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 33/5038 : involving detection of metabolites per se
• CPC Class Codes: G01N 35/00584 : Control arrangements for automatic analysers
• CPC Class Codes: G01N 35/1002 : Reagent dispensers

Abstract

Devices and methods that measure one or more properties of a living cell culture that is contained in liquid media within a vessel, and typically analyzes plural cell cultures contained in plural vessels such as the wells of a multiwell microplate substantially in parallel. The devices incorporate a sensor that remains in equilibrium with, e.g., remains submerged within, the liquid cell media during the performance of a measurement and during addition of one or more cell affecting fluids such as solutions of potential drug compounds.

Claims

1. A cartridge which mates with a plate defining a plurality of wells, the cartridge comprising:

a substantially planar element comprising

a plurality of regions corresponding to a number of respective openings of the wells in the plate, the regions being spaced apart so as to match a spacing of at least a portion of the multiple wells;

located in plural respective regions of the cartridge, at least one of

a sensor to analyze a constituent in a well, and

an aperture to receive a sensor, and

at least one port formed in the cartridge in at least a subset of the regions, the port being adapted to deliver a test fluid to a respective well of the plate during the time said sensor is disposed within liquid within said well.

2. The cartridge of claim 1, wherein the port forms a capillary aperture to retain test fluid in the port absent an external force.

3. The cartridge of claim 2, wherein the external force is selected from the group consisting of a positive pressure differential force, a negative pressure differential force, and a centrifugal force.

4. The cartridge of claim 1, wherein the sensor is compliantly attached to the planar element.

5. The cartridge of claim 1, further comprising a second port formed in the cartridge in the at least one region, the second port adapted to deliver a second test fluid to the respective well.

6. The cartridge of claim 1, wherein the cartridge forms a cover for the multiwell plate to reduce at least one of contamination and evaporation of sample in the multiwell plate.

7. The cartridge of claim 1, further comprising, in multiple regions of the cartridge, at least one port and at least one of the sensor and an aperture adapted to receive the sensor.

8. The cartridge of claim 7, wherein all of the ports are in fluidic communication.

9. The cartridge of claim 7, further comprising a second port formed in the cartridge in every region.

10. The cartridge of claim 9, wherein the second ports are in fluidic communication with each other and not in fluidic communication with other ports.

11. The cartridge of claim 7 comprising 24, 96, or 384 regions mated with 24, 96, or 384 well multiwell plates, each region comprising said sensor or a portion thereof or an aperture adapted to receive the sensor, and a set of 2, 3, 4, 5, 6, 7, or 8 ports.

12. The cartridge of claim 11, further comprising a multiplexer in fluidic communication with each set of ports.

13. The cartridge of claim 12, wherein the multiplexer is adapted to connect to a single pneumatic source to permit delivering fluid to the wells sequentially from each set of ports.

14. The cartridge of claim 1 wherein the at least one of the sensor or portion thereof adapted to analyze a constituent in a well and the aperture adapted to receive the sensor comprises a sensor sleeve structure having a light translucent surface and disposed thereon a fluorophore having fluorescent properties dependent on at least one of the presence and the concentration of a constituent in the well.

15. The cartridge of claim 14 wherein the sensor sleeve further comprises an elongate housing to receive a wave guide for at least one of stimulating the fluorophore and receiving fluorescent emissions from the fluorophore.