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Diving under a Microscope—A New Simple and Versatile In Vitro Diving Device for Fluorescence and Confocal Microscopy Allowing the Controls of Hydrostatic Pressure, Gas Pressures, and Kinetics of Gas Saturation

Published online by Cambridge University Press:  17 April 2013

Qiong Wang ,
Marc Belhomme ,
François Guerrero ,
Aleksandra Mazur ,
Kate Lambrechts  and
Michaël Theron
Qiong Wang
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
Marc Belhomme
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
François Guerrero
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
Aleksandra Mazur
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
Kate Lambrechts
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
Michaël Theron*
Affiliation:
Laboratory ORPHY, Université Européenne de Bretagne, Université de Brest, 6 Avenue Le Gorgeu CS 93837, 29238 Brest-CEDEX 3, France
*
*Corresponding author. E-mail: Michael.theron@univ-brest.fr
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Abstract

How underwater diving effects the function of the arterial wall and the activities of endothelial cells is the focus of recent studies on decompression sickness. Here we describe an in vitro diving system constructed to achieve real-time monitoring of cell activity during simulated dives under fluorescent microscopy and confocal microscopy. A 1-mL chamber with sapphire windows on both sides and located on the stage of an inverted microscope was built to allow in vitro diving simulation of isolated cells or arteries in which activities during diving are monitored in real-time via fluorescent microscopy and confocal microscopy. Speed of compression and decompression can range from 20 to 2000 kPa/min, allowing systemic pressure to range up to 6500 kPa. Diving temperature is controlled at 37°C. During air dive simulation oxygen partial pressure is optically monitored. Perfusion speed can range from 0.05 to 10 mL/min. The system can support physiological viability of in vitro samples for real-time monitoring of cellular activity during diving. It allows regulations of pressure, speeds of compression and decompression, temperature, gas saturation, and perfusion speed. It will be a valuable tool for hyperbaric research.

Keywords

air divedecompression sicknessactivity of arterial endothelial cellkinetics of gas saturationin vitro real-time monitoringfluorescence and confocal microscopy
Type
Equipment and Techniques Development: Biological
Information
Microscopy and Microanalysis , Volume 19 , Issue 3 , June 2013 , pp. 608 - 616
Copyright
Copyright © Microscopy Society of America 2013 

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