A multipurpose modular system for high-resolution microscopy at high hydrostatic pressure.

We have developed a modular system for high-resolution microscopy at high hydrostatic pressure. The system consists of a pressurized cell of volume approximately 100 microl, a temperature controlled holder, a ram, and a piston. We have made each of these components in several versions which can be interchanged to allow a wide range of applications. Here, we report two pressure cells with pressure ranges 0.1-700 MPa and 0.1-100 MPa, which can be combined with hollow or solid rams and pistons. Our system is designed to work with fluorescent samples (using a confocal or epifluorescence microscope), but also allows for transmitted light microscopy via the hollow ram and piston. The system allows precise control of pressure and temperature (-20 to 70 degrees C), as well as rapid pressure quenching. We demonstrate its performance and versatility with two applications: time-resolved imaging of colloidal phase transitions caused by pressure changes between 0.1 and 100 MPa, and imaging the growth of Escherichia coli bacteria at 50 MPa. We also show that the isotropic-nematic phase transition of pentyl-cyanobiphenyl (5CB) liquid crystal provides a simple, convenient, and accurate method for calibrating pressure in the range 0.1-200 MPa.