Preparation and characterization of sensing layers for pH detection in living plant cells

J. Mrazek; I. Kasik; J. Nekola; T. Martan; O. Podrazky; M. Pospisilova; V. Matejec

doi:10.1117/12.886250

6 May 2011 Preparation and characterization of sensing layers for pH detection in living plant cells

J. Mrazek, I. Kasik, J. Nekola, T. Martan, O. Podrazky, M. Pospisilova, V. Matejec

Proceedings Volume 8073, Optical Sensors 2011; and Photonic Crystal Fibers V; 807318 (2011) https://doi.org/10.1117/12.886250
Event: SPIE Optics + Optoelectronics, 2011, Prague, Czech Republic

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.

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J. Mrazek, I. Kasik, J. Nekola, T. Martan, O. Podrazky, M. Pospisilova, and V. Matejec "Preparation and characterization of sensing layers for pH detection in living plant cells", Proc. SPIE 8073, Optical Sensors 2011; and Photonic Crystal Fibers V, 807318 (6 May 2011); https://doi.org/10.1117/12.886250

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KEYWORDS

Luminescence

Absorption

Transducers

Sensors

Matrices

Calibration

Biological and chemical sensing

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