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High pressure sample cell for total internal reflection fluorescence spectroscopy at pressures up to 2500 bar.
MedLine Citation:
PMID:  22938334     Owner:  NLM     Status:  In-Data-Review    
Total internal reflection fluorescence (TIRF) spectroscopy is a surface sensitive technique that is widely used to characterize the structure and dynamics of molecules at planar liquid-solid interfaces. In particular, biomolecular systems, such as protein adsorbates and lipid membranes can easily be studied by TIRF spectroscopy. Applying pressure to molecular systems offers access to all kinds of volume changes occurring during assembly of molecules, phase transitions, and chemical reactions. So far, most of these volume changes have been characterized in bulk solution, only. Here, we describe the design and performance of a high pressure sample cell that allows for TIRF spectroscopy under high pressures up to 2500 bar (2.5 × 10(8) Pa), in order to expand the understanding of volume effects from the bulk phase to liquid-solid interfaces. The new sample cell is based on a cylindrical body made of Nimonic 90 alloy and incorporates a pressure transmitting sample cuvette. This cuvette is composed of a fused silica prism and a flexible rubber gasket. It contains the sample solution and ensures a complete separation of the sample from the liquid pressure medium. The sample solution is in contact with the inner wall of the prism forming the interface under study, where fluorescent molecules are immobilized. In this way, the new high pressure TIRF sample cell is very useful for studying any biomolecular layer that can be deposited at a planar water-silica interface. As examples, high pressure TIRF data of adsorbed lysozyme and two phospholipid membranes are presented.
Juny Koo; Claus Czeslik
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Review of scientific instruments     Volume:  83     ISSN:  1089-7623     ISO Abbreviation:  Rev Sci Instrum     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-09-03     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0405571     Medline TA:  Rev Sci Instrum     Country:  United States    
Other Details:
Languages:  eng     Pagination:  085109     Citation Subset:  IM    
Technische Universität Dortmund, Fakultät Chemie, D-44221 Dortmund, Germany.
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