Document Detail


A noninvasive thin film sensor for monitoring oxygen tension during in vitro cell culture.
MedLine Citation:
PMID:  19860390     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Oxygen tension in mammalian cell culture can profoundly affect cellular differentiation, viability, and proliferation. However, precise measurement of dissolved oxygen in real time remains difficult. We report a new noninvasive sensor that can accurately measure oxygen concentration during cell culture while being compatible with live-cell imaging techniques such as fluorescence and phase contrast microscopy. The sensor is prepared by integrating the porphyrin dye, Pt(II) meso-tetrakis(pentafluorophenyl)porphine (PtTFPP) into polydimethylsiloxane (PDMS) thin films. Response of the sensor in the presence of oxygen can be characterized by the linear Stern-Volmer relationship with high sensitivity (K(SV) = 584 +/- 71 atm(-1)). A multilayer sensor design, created by sandwiching the PtTFPP-PDMS with a layer of Teflon AF followed by a second PDMS layer, effectively mitigates against dye cytotoxicity while providing a substrate for cell attachment. Using this sensor, changes in oxygen tension could be monitored in real-time as attached cells proliferated. The oxygen tension was found to decrease due to oxygen consumption by the cells, and the data could be analyzed using Fick's law to obtain the per-cell oxygen consumption rate. This sensor is likely to enable new studies on the effects of dissolved oxygen on cellular behavior.
Authors:
Peter C Thomas; Michael Halter; Alessandro Tona; Srinivasa R Raghavan; Anne L Plant; Samuel P Forry
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical chemistry     Volume:  81     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2009 Nov 
Date Detail:
Created Date:  2009-11-13     Completed Date:  2010-02-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  9239-46     Citation Subset:  IM    
Affiliation:
Biochemical Science Division, National Institutes of Standard and Technology, Gaithersburg, Maryland 20899, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques / methods*
Cell Survival / drug effects
Dimethylpolysiloxanes / chemistry*
Mice
NIH 3T3 Cells
Oxygen / analysis*
Porphyrins / chemistry*
Chemical
Reg. No./Substance:
0/Dimethylpolysiloxanes; 0/Porphyrins; 63148-62-9/baysilon; 7782-44-7/Oxygen

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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