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Proceedings Paper

Preparation and characterization of sensing layers for pH detection in living plant cells
Author(s): J. Mrazek; I. Kasik; J. Nekola; T. Martan; O. Podrazky; M. Pospisilova; V. Matejec
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Paper Abstract

Local chemical sensing in living cells by fluorescence methods with submicron spatial resolution is in the scope of biologist because of bringing new information about biochemical processes in living matter [1]. One of the most important monitored variables is pH. Despite of progress of novel submicron probes suitable for in-situ measurement in living cells [1] and biological micro samples [2] still there is a lack off suitable opto-chemical transducers sensitive around pH 5-7 limiting development of novel fluorescence sensors. Moreover, the interaction of the immobilized transducer with the matrix can strongly affect its fluorescence properties. In our contributition the 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) fluorescence pH transducer was incorporated into organosols based on tetraethylorthosilicate (TEOS), 3-glycidoxypropyltrimethoxysilane (GPTMS) and 3-aminopropyltriethoxysilane (APTES). Formed organosols were spin-coated onto Pyrex glass substrates and thermally treated at 140°C for 4 hours. Prepared thin layers were exposed to Britton-Robinson buffers with different pH ranging from 4 to 8 pH units. Optical properties of immobilized BCECF were investigated by the mean of absorption and fluorescence spectroscopy Acquired results were compared with the properties of BCECF solutions. It was found that all matrices reduce the sensitivity of the BCECF transducer comparing to the free solution. GPTMS and APTES contained layers exhibited better mechanical properties and increase the solubility of BCECF inside prepared layers comparing to layers prepared from pure TEOS.

Paper Details

Date Published: 7 May 2011
PDF: 6 pages
Proc. SPIE 8073, Optical Sensors 2011; and Photonic Crystal Fibers V, 807318 (7 May 2011); doi: 10.1117/12.886250
Show Author Affiliations
J. Mrazek, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
I. Kasik, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
J. Nekola, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
T. Martan, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
O. Podrazky, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
M. Pospisilova, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)
V. Matejec, Institute of Photonics and Electronics of the ASCR, v.v.i. (Czech Republic)


Published in SPIE Proceedings Vol. 8073:
Optical Sensors 2011; and Photonic Crystal Fibers V
Kyriacos Kalli; Francesco Baldini; Jiri Homola; Robert A. Lieberman, Editor(s)

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