ABSTRACT Oxygen consumption is a fundamental component of metabolic networks, mitochondrial function, and global carbon cycling. To date there is no method available that allows for replicate measurements on attached and unattached biological samples without compensation for extraneous oxygen leaking into the system. Here we present the Respiratory Detection System, which is compatible with virtually any biological sample. The RDS can be used to measure oxygen uptake in microliter-scale volumes with a reversibly sealed sample chamber, which contains a porphyrin-based oxygen sensor. With the RDS, one can maintain a diffusional seal for up to three hours, allowing for the direct measurement of respiratory function of samples with fast or slow metabolic rates. The ability to easily measure oxygen uptake in small volumes with small populations or dilute samples has implications in cell biology, environmental biology, and clinical diagnostics.
Cite this paper
nullStrovas, T. , McQuaide, S. , Anderson, J. , Nandakumar, V. , Kalyuzhnaya, M. , Burgess, L. , Holl, M. , Meldrum, D. and Lidstrom, M. (2010) Direct measurement of oxygen consumption rates from attached and unattached cells in a reversibly sealed, diffusionally isolated sample chamber. Advances in Bioscience and Biotechnology, 1, 398-408. doi: 10.4236/abb.2010.15053.
 Strovas, T.J., Dragavon, J.M., Hankins, T.J., Callis, J.B., Burgess, L.W. and Lidstrom, M.E. (2006) Measurement of respiration rates of Methylobacterium extorquens AM1 cultures by use of a phosphorescence-based sensor. Applied Environmental Microbiology, 72, 1692-1695.
Bidanda, B. (2005) Respiration in aquatic ecosystems. In: Del Giorgio, P.A. and Williams, P.J. le B., Eds., Journal of Plankton Research, Oxford University Press, New York.
Fink, S.L. and Cookson, B.T. (2005) Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells. Infection and Immunity, 73, 1907-1916.
Weissig, V., Boddapati, S.V., Jabr L., and D'Souza, G.G. (2007) Mitochondria-specific nanotechnology. Nanomedicine, 2, 275-285.
Wu, M., Neilson, A., Swift, A.L., Moran, R., Tamagnine, J., Parslow, D., Armistead, S., Lemire, K., Orrell, J., Teich, J., Chomicz, S. and Ferrick, D.A. (2007) Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. American Journal of Physiology: Cellular Physiology, 292, 125-136.
Bell, E.L. andChandel, N.S. (2007) Mitochondrial oxygen sensing: Regulation of hypoxia-inducible factor by mitochondrial generated reactive oxygen species. Essays in Biochemistry, 43, 17-27.
Fink, S.L. and Cookson, B.T. (2007) Pyroptosis and host cell death responses during Salmonella infection. Cellular Microbiology, 9, 2562-2570.
Maley, C.C. (2007) Multistage carcinogenesis in Barrett’s esophagus. Cancer Letters, 245, 22-32.
Eklund, S.E., Thompson, R.G., Snider, R.M., Carney, C. K., Wright, D.W., Wikswo, J. and Cliffel, D.E. (2009) Metabolic discrimination of select list agent by monitoring cellular responses in a multianalyte microphysiometer. Sensors, 9, 2117-2133.
Ramamoorthy, R., Dutta, P.K. and Akbar, S.A. (2003) Oxygen sensors: Materials, methods, designs, and applications. Journal of Material Science, 38, 4271-4282.
Renger, G. and Hanssum, B. (2009) Oxygen detection in biological systems. Photosynthesis Research, 102, 487-498.
Kuang, Y. and Walt, D.R. (2007) Detecting oxygen consumption in the proximity of Saccharomyces cerevisiae cells using self-assembled fluorescent nanosensors. Biotechnology Bioengineering, 96, 318-325.
Saito, T., Wu, C.C., Shiku, H., Yasukawa, T., Yokoo, M., Ito-Sasaki, T., Abe, H., Hoshi, H. and Matsue, T. (2006) Oxygen consumption of cell suspension in a poly (dimethylsiloxane) (PDMS) microchannel estimated by scanning electrochemical microscop. Analyst, 131, 1006-1011.
Warkentin, M., Freese, H.M., Karsten, U. and Schumann, R. (2007) New and fast method to quantify respiration rates of bacterial and plankton communities in freshwater ecosystems by using optical oxygen sensor spots. Applied Environmental Microbiology, 73, 6722-6729.
Papkovsky, D.B. and O’Riordan, T.C. (2007) Emerging applications of phosphorescent metalloporphyrins. Journal of Fluorescence, 15, 569-584.
Abe, H. (2007) A non-invasive and sensitive method for measuring cellular respiration with a scanning electrochemical microscopy to evaluate embryo quality. Journal of Mammalian Ova Research, 24, 70-78.
Borisov, S.M., Nuss, G. and Klimant, I. (2008) Red light-excitable oxygen sensing materials based on platinum(II) and palladium(II) benzoporphyrins. Analytical Chemistry, 80, 9435-9442.
Molter, T.W., McQuaide, S.C., Suchorolski, M.T., Strovas, T.J., Burgess, L.W., Meldrum, D.R. and Lidstrom, M.E. (2009) A microwell array device capable of measuring single-cell-oxygen consumption rates. IEEE Sensors and Actuators B, 135, 678-686.
O’Donovan, C., Twomey, E., Alderman, J., Moore, T. and Papkovsky, D. (2006) Development of a respirometric biochip for embryo assessment. Lab on a Chip, 6, 1438-1444.
O'Mahony, F.C., O’Donovan, C., Hynes, J., Moore, T., Davenport, J. and Papkovsky, D.B. (2005) Optical oxygen microrespirometry as a platform for environmental toxicology and animal model studies. Environmental Science and Technology, 39, 5010-5014.
Wilson, D.F., Vanderkooi, J.M., Green, T.J., Maniara, G., Defeo, S.P. and Bloomgarden, D.C. (1987) A versatile and sensitive method for measuring oxygen. Advances in Experimental Medical Biology, 215, 71-77.
Alderman, J., Hynes, J., Floyd, S.M., Krüger, J., O’Conner, R. and Papkovsky, D.B. (2004) A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing. Biosensors and Bioelectronics, 19, 1529-1535.
Birmele, M., Roberts, M. and Garland, J. (2006) Characterization of methods for determining sterilization efficacy and reuse efficiency of oxygen biosensor multiwell plates. Journal of Microbiological Methods, 67, 619-623.
Dragavon, J., Molter, T., Young, C., Strovas, T., McQuaide, S., Holl, M., Zhang, M., Cookson, B., Jen, A., Lidstrom, M. and Burgess, L. (2008) A cellular isolation system for real-time single-cell oxygen consumption monitoring. Journal of the Royal Society Interface, 5(S2), S151-S159.
Will, Y., Hynes, J., Ogurtsov, V.I. and Papkovsky, D.B. (2007) Analysis of mitochondrial function using phosphorescent oxygen-sensitive probes. Nature Protocols, 1, 2563-2572.
Wodnicka, M., Guarino, R.D., Hemperly, J.J., Timmins, M.R., Stitt, D. and Pitner, J.B. (2000) Novel fluorescent technology platform for high throughput cytotoxicity and proliferation assays. Journal of Biomolecular Screening, 5, 141-152.
Arain, S., Weiss, S., Heinzle, E., John, G.T., Krause, C. and Klimant, I. (2005) Gas sensing in microplates with optodes: Influence of oxygen exchange between sample, air, and plate material. Biotechnology and Bioengineering, 90, 271-280.
Gerencser, A.A., Neilson, A., Choi, S.W., Edman, U., Yadava, N., Oh, R.J., Ferrick, D.A., Nicholls, D.G. and Brand, M.D. (2009) Quantitative microplate-based respirometry with correction for oxygen diffusion. Analytical Chemistry, 81, 6868-6878.
Strovas, T.J., Sauter, L.M., Guo, X. and Lidstrom, M.E. (2007) Cell-to-cell heterogeneity in growth rate and gene expression in Methylobacterium extorquens AM1. Journal of Bacteriology, 189, 7127-7133.
Kalyuzhnaya, M.G., Bowerman, S., Nercessian, O., Lidstrom, M.E. and Chistoserdova, L. (2005) Highly divergent genes for methanopterin-linked C1 transfer reactions in Lake Washington, assessed via metagenomic analysis and mRNA detection. Applied Environmental Microbiology, 71, 8846-8854.
Guo, X. (2006) Intergrative study of metabolism of Methylobacterium extorquens AM1 from a metabolomic perspective. Ph.D. dissertation, University of Washington, Department of Chemical Engineering.
Abramoff, M.D., Magelhaes, P.J. and Ram, S.J. (2004) Image processing with ImageJ. Biophotonics International, 11, 36-42.
Carpenter, A.E., Jones, T.R., Lamprecht, M.R., Clarke, C. Kang, I.H., Friman, O., Guertin, D.A., Chang, J.H., Lindquist, R.A., Moffat, J., Golland, P. and Sabatini, D. M. (2006) CellProfiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biology, 7, R100.
Nalwaya, N. and Deen, W.M. (2005) Nitric oxide, oxygen, and superoxide formation and consumption in macrophage cultures. Chemical Research in Toxicology, 18, 486-93.
Leschelle, X., Goubern, M., Andriemihaja, M., Blottiere, H.M., Coupland, E., Gonzalez-Barroso, M.D., Petite, C., Pagniez, A., Chaumentet, C., Mignotte, B., Bouillaud, F. and Blachier, F. (2005) Adaptative metabolic response of human colonic epithelial cells to the adverse effects of the luminal compound sulfide. Biochimica et Biophysica Acta, 1725, 201-212.
Heerlein, K., Schulze, A., Hotz, L., Bartsch, P. and Mairbaurl, H. (2005) Hypoxia decreases cellular ATP demand and inhibits mitochondrial respiration of A549 cells. American Journal of Respiratory Cell and Molecular Biology, 32, 44-51.
Lidstrom, M.E. and Meldrum, D.R. (2003) Life-on-a-chip. Nature Reviews Microbiology, 1, 158-164.