A cellular isolation system for real-time single-cell oxygen consumption monitoring.
The development of a cellular isolation system (CIS) that enables the monitoring of single-cell oxygen consumption rates in real time is presented. The CIS was developed through a multidisciplinary effort within the Microscale Life Sciences Center (MLSC) at the University of Washington. The system comprises arrays of microwells containing Pt-porphyrin-embedded polystyrene microspheres as the reporter chemistry, a lid actuator system and a gated intensified imaging camera, all mounted on a temperature-stabilized confocal microscope platform. Oxygen consumption determination experiments were performed on RAW264.7 mouse macrophage cells as proof of principle. Repeatable and consistent measurements indicate that the oxygen measurements did not adversely affect the physiological state of the cells measured. The observation of physiological rates in real time allows studies of cell-to-cell heterogeneity in oxygen consumption rate to be performed. Such studies have implications in understanding the role of mitochondrial function in the progression of inflammatory-based diseases, and in diagnosing and treating such diseases.
Department of Chemistry, University of Washington, Seattle, WA 98195-2180, USA. joedragavon@gmail.com
Joe
J
Dragavon
Tim
T
Molter
Cody
C
Young
Tim
T
Strovas
Sarah
S
McQuaide
Mark
M
Holl
Meng
M
Zhang
Brad
B
Cookson
Alex
A
Jen
Mary
M
Lidstrom
Deirdre
D
Meldrum
Lloyd
L
Burgess
D000818:Animals; D002138:Calibration; D019069:Cell Respiration; D002469:Cell Separation / Q000295:instrumentation* / Q000592:standards; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000295:instrumentation; D051379:Mice; D010101:Oxygen Consumption; D015203:Reproducibility of Results