Device for monitoring cells

• US 20050191613A1
• Filed: 10/04/2004
• Published: 09/01/2005
• Est. Priority Date: 04/18/1991
• Status: Active Grant

• Current Assignee: David T. Stitt, Gregory J. Burrell, John Jacob Hemperly, Magdalena Wodnicka, Patrick Shawn Beaty, Pitner J. Bruce, Richard D. Guarino, Timothy G. Foley Jr
• Original Assignee: David T. Stitt, Gregory J. Burrell, John Jacob Hemperly, Magdalena Wodnicka, Patrick Shawn Beaty, Pitner J. Bruce, Richard D. Guarino, Timothy G. Foley Jr
• Inventors: Beaty, Patrick Shawn, Pitner, J. Bruce, Burrell, Gregory J., Guarino, Richard D., Foley, Timothy G. Jr., Stitt, David T., Hemperly, John Jacob, Wodnicka, Magdalena
• Granted Patent: US 7,338,778 B2
• Time in Patent Office: Days
• US Class Current: 435/4
• 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

1-96. -96. (canceled)

97. A method for analyzing mammalian cells based on their consumption of oxygen over time, the method comprising:

(i) providing a sample comprising the cells;

(ii) placing the sample in chemical communication with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property when irradiated with light containing wavelengths which cause the compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is nondestructive to the cells;

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

(iv) measuring or visually observing the change in luminescent property from the luminescent compound while irradiating the sensor compound with the light, over a period of time.

98. The method of claim 97, further comprising:

(v) comparing the measurement of step (iv) to a control.

99. The method of claim 97 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.

100. The method of claim 99, wherein the matrix is a rubber or plastic matrix.

101. The method of claim 99, wherein the matrix is a silicone rubber matrix.

102. The method of claim 97, wherein the luminescent compound is adsorbed on solid silica particles.

103. The method of claim 97, wherein the luminescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) salt.

104. The method of claim 103, wherein the luminescent compound is tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.

105. The method of claim 97, wherein the luminescent compound is a tris-2,2′

-bipyridyl ruthenium (II) salt.

106. The method of claim 105, wherein the luminescent compound is tris-2,2′

-bipyridyl ruthenium (II) chloride hexahydrate.

107. The method of claim 97, wherein the luminescent compound is 9,10-diphenyl anthracene.

108. The method of claim 97, wherein the sample is exposed to atmospheric oxygen.

109. The method of claim 97 wherein, in step (i), the sample is also contacted with an effective concentration of one or more biomaterials.

110. The method of claim 109, wherein the biomaterial is MATRIGEL®

.

111. The method of claim 97, wherein, the sample is also contacted with an effective concentration of one or more extracellular matrices.

112. The method of claim 111, wherein the extracellular matrix is collagen.

113. The method of claim 97, wherein the sample is contacted with an effective concentration of one or more additives or coating substances.

114. The method of claim 97, wherein the period of time is approximately 6 hours or less.