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Capillary-based, high-pressure chamber for fluorescence microscopy imaging

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4 Author(s)
Raber, Erica C. ; Department of Physics, Miami University, Oxford, Ohio 45056 ; Dudley, Jonathan A. ; Salerno, Michael ; Urayama, Paul

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We present a high-pressure chamber suitable for the fluorescence microscopy imaging of cellular systems, consisting of a quartz capillary attached to high-pressure tubing with the capillary walls acting as the optical window and mechanical support. The chamber withstands pressures up to 1000 atm, spanning most of the biosphere’s pressure range. We validate the chamber for high-pressure studies by using a pH-sensitive fluorophore to sense the pressure-induced acidification of a phosphate buffer. Next, because the capillary walls result in image aberrations, we demonstrate the ability to use spatial deconvolution to improve image resolution. Capillary wall thicknesses are less than 0.5 mm, making the chamber compatible with high-numerical-aperture optics.

Published in:

Review of Scientific Instruments  (Volume:77 ,  Issue: 9 )