Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads
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2013-04-21
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Lin Wang, Miguel A. Acosta, Jennie B. Leach and Rebecca L. Carrier, Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads , Lab Chip. 2013 Apr 21; 13(8): 1586–1592. doi: 10.1039/c3lc41366g
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Abstract
Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of
microns scale) is often desirable but difficult to achieve in cell culture. In this study,
biocompatible oxygen sensing beads were prepared and tested for their potential for real-time
monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems.
Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive
Ru(Ph₂phen₃)Cl₂ dye and oxygen insensitive Nile blue reference dye, and a poly-dimethylsiloxane
(PDMS) shell rendering biocompatibility. Human intestinal epithelial Caco-2 cells were cultivated
on a series of PDMS and type I collagen based substrates patterned with micro-well arrays for 3 or
7 days, and then brought into contact with oxygen sensing beads. Using an image analysis
algorithm to convert florescence intensity of beads to partial oxygen pressure in the culture
system, tens of microns-size oxygen sensing beads enabled the spatial measurement of local
oxygen concentration in the microfabricated system. Results generally indicated lower oxygen
level inside wells than on top of wells, and local oxygen level dependence on structural features of
cell culture surfaces. Interestingly, chemical composition of cell culture substrates also appeared
to affect oxygen level, with type-I collagen based cell culture systems having lower oxygen
concentration compared to PDMS based cell culture systems. In general, results suggest that
oxygen sensing beads can be utilized to achieve real-time and local monitoring of microenvironment
oxygen level in 3D microfabricated cell culture systems.