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Microscopy under pressure—An optical chamber system for fluorescence microscopic analysis of living cells under high hydrostatic pressure
Author(s) -
Frey Benjamin,
Hartmann Markus,
Herrmann Martin,
MeyerPittroff Roland,
Sommer Karl,
Bluemelhuber Gerrit
Publication year - 2006
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20269
Subject(s) - hydrostatic pressure , propidium iodide , fluorescence microscope , fluorescence , optical microscope , microscopy , microscope , bright field microscopy , phase contrast microscopy , chemistry , biophysics , live cell imaging , materials science , optics , nanotechnology , cell , scanning electron microscope , biology , composite material , physics , biochemistry , apoptosis , programmed cell death , thermodynamics
High hydrostatic pressure (HHP) becomes more and more interesting for life science research, since it can be employed to inactivate various cells. To directly monitor “cells under pressure,” the development of an optical high‐pressure chamber is required. Therefore, an optical pressure chamber that can be used for up to 300 MPa was constructed. This chamber has already been described as a tool for in situ observation of dynamic changes of microscopic structures in bright field as well as phase contrast. In combination with an inverted microscope, we obtained brilliant microscopic color pictures with an optical resolution more than 0.56 μm. Here, we demonstrate the capabilities of the HHP cell, in combination with epifluorescence microscopy. Using a nonadherent human B‐cell line (Raji, ATCC CCL 86), stained with the fluorescent dyes propidium iodide, Hoechst 33342, or dihexyloxacarbocyanine iodide, we were able to show that the system is suitable to perform fluorescence microscopic analyses, with pressures up to 300 MPa, with viable mammalian cells. Microsc. Res. Tech. 69:65–72, 2006. © 2006 Wiley‐Liss, Inc.