A perfusion system for the fluorescence-based monitoring of physiological responses to high hydrostatic pressures

POSTER

Abstract

Hydrostatic pressures of a few-hundred atmospheres affect biological components and processes, for example, the stability of macromolecular assemblies, the biophysical properties of membranes, enzymatic activity, and protein structure. These in turn lead to a range of pressure effects on cellular physiology. Here we present a mirco-perfusion system designed for the real-time spectroscopic monitoring of cellular systems under high pressure. The system consists of an optically-compatible pressure chamber and interchangeable fluid reservoirs. Perfusion is achieved using a dual pressure-generator configuration, where one positive-displacement generator is compressed while the other is retracted, thus maintaining pressure while achieve fluid flow. Control over perfusion rate (typically in the 10 $\mu $l/s range) and the ability to change between fluid reservoirs while under pressure (up to 600 atm) is demonstrated. The real-time monitoring of the response of cellular autofluorescence to mitochondrial functional modifiers (e.g., cyanide) while under pressure is also demonstrated.

Authors

  • Paul Urayama

    Miami University

  • John Royston

    Ball State University, Naval Research Laboratory, Washington, DC 20375, USA, Univ of Cincinnati, KITP China, U.C. Berkeley, FNAL, Cornell, West Virginia University, University of Pittsburgh, The Ohio State University, Carnegie Mellon University, Miami University, University of Notre Dame, University of Nebraska-Lincoln, Miami Univ, Australia National Univ., Miami Univ., Univ. of Cincinnati, Physics and Astronomy Department, Ohio University, Athens, OH 45701, Australian National University, University of Toledo, The University of Toledo, University of Toledo, Wright Center for Photovoltaics Innovation and Commercialization, University of Cincinnati, University of California, Davis

  • John Royston

    Ball State University, Naval Research Laboratory, Washington, DC 20375, USA, Univ of Cincinnati, KITP China, U.C. Berkeley, FNAL, Cornell, West Virginia University, University of Pittsburgh, The Ohio State University, Carnegie Mellon University, Miami University, University of Notre Dame, University of Nebraska-Lincoln, Miami Univ, Australia National Univ., Miami Univ., Univ. of Cincinnati, Physics and Astronomy Department, Ohio University, Athens, OH 45701, Australian National University, University of Toledo, The University of Toledo, University of Toledo, Wright Center for Photovoltaics Innovation and Commercialization, University of Cincinnati, University of California, Davis

  • John Royston

    Ball State University, Naval Research Laboratory, Washington, DC 20375, USA, Univ of Cincinnati, KITP China, U.C. Berkeley, FNAL, Cornell, West Virginia University, University of Pittsburgh, The Ohio State University, Carnegie Mellon University, Miami University, University of Notre Dame, University of Nebraska-Lincoln, Miami Univ, Australia National Univ., Miami Univ., Univ. of Cincinnati, Physics and Astronomy Department, Ohio University, Athens, OH 45701, Australian National University, University of Toledo, The University of Toledo, University of Toledo, Wright Center for Photovoltaics Innovation and Commercialization, University of Cincinnati, University of California, Davis