Luminescent method for determining cell oxygen consumption

• US 7,338,778 B2
• Filed: 10/04/2004
• Issued: 03/04/2008
• Est. Priority Date: 04/18/1991
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Beaty, Patrick Shawn, Burrell, Gregory J., Guarino, Richard D., Foley, Timothy G. Jr., Stitt, David T., Hemperly, John Jacob, Wodnicka, Magdalena, Pitner, J. Bruce
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US10/957,802
• Publication Number:
  US 20050191613A1
• Time in Patent Office: 1,247 Days
• Field of Search: 435/34, 435/29, 435/69.1, 435/32
• US Class Current: 435/34
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/18 Testing for antimicrobial activity of a material
• CPC Class Codes: C12Q 2304/44 Oxygen
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

The present invention relates to methods for detection and evaluation of metabolic activity of eukaryotic and/or prokaryotic cells based upon their ability to consume dissolved oxygen. The methods utilize a luminescence detection system which makes use of the sensitivity of the luminescent emission of certain compounds to the presence of oxygen, which quenches (diminishes) the compound'"'"'s luminescent emission in a concentration dependent manner. Respiring eukaryotic and/or prokaryotic cells will affect the oxygen concentration of a liquid medium in which they are immersed. Thus, this invention provides a convenient system to gather information on the presence, identification, quantification and cytotoxic activity of eukaryotic and/or prokaryotic cells by determining their effect on the oxygen concentration of the media in which they are present.

Claims

1. A method for determining oxygen consumption by cells comprising the steps of:

providing a sample comprising a mammalian cell culture;

placing the sample in chemical communication with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property, upon exposure to oxygen, wherein the presence of the sensor composition is nondestructive to the mammalian cells;

irradiating the sensor composition with light containing wavelengths which cause the luminescent compound to luminescence; and

determining the change in luminescent property, over a period of time, wherein the change in luminescent property is indicative of a change in oxygen consumption by the mammalian cells.

2. The method of claim 1, further comprising the step of:

comparing the determined change in luminescent property to a control.

3. The method of claim 1 wherein the luminescent compound is contained within a matrix that is relatively impermeable to water and non-gaseous solutes, but which has a high permeability to oxygen.

4. The method of claim 3, wherein the matrix is a rubber or plastic matrix.

5. The method of claim 3, wherein the matrix is a silicone rubber matrix.

6. The method of claim 1, wherein the luminescent compound is adsorbed on solid silica particles.

7. The method of claim 1, wherein the luminescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) salt.

8. The method of claim 7, wherein the luminescent compound is tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.

9. The method of claim 1, wherein the luminescent compound is a tris-2,2′

-bipyridyl ruthenium (II) salt.

10. The method of claim 9, wherein the luminescent compound is tris-2,2′

-bipyridyl ruthenium (II) chloride hexahydrate.

11. The method of claim 1, wherein the luminescent compound is 9,10-diphenyl anthracene.

12. The method of claim 1, wherein the sample is exposed to atmospheric oxygen.

13. The method of claim 1, wherein, the sample is also contacted with one or more extracellular matrices.

14. The method of claim 13, wherein the extracellular matrix is collagen.

15. The method of claim 1, wherein the period of time is not greater than approximately 6 hours.