In vivo optical detection of pH in microscopic tissue samples of Arabidopsis thaliana

0424363 - ÚFE 2014 RIV NL eng J - Journal Article
Kašík, Ivan - Podrazký, Ondřej - Mrázek, Jan - Martan, Tomáš - Matějec, Vlastimil - Hoyerová, Klára - Kamínek, Miroslav
In vivo optical detection of pH in microscopic tissue samples of Arabidopsis thaliana.
Materials Science & Engineering C-Materials for Biological Applications. Roč. 33, č. 8 (2013), s. 4809-4815. ISSN 0928-4931. E-ISSN 1873-0191
R&D Projects: GA ČR GAP102/10/2139
Institutional support: RVO:67985882 ; RVO:61389030
Keywords : Ratiometric fluorescence * Arabidopsis thaliana * Tissue
Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering
Impact factor: 2.736, year: 2013

Minimally invasive in vivo measurement of pH inmicroscopic biological samples of μmor μl size, e.g. plant cells, tissues and saps, may help to explain complex biological processes. Consequently, techniques to achieve such measurements are a focus of interest for botanists. This paper describes a technique for the in vivo measurement of pH in the range pH 5.0 to pH 7.8 in microscopic plant tissue samples of Arabidopsis thaliana based on a ratiometric fluorescence method using low-loss robust tapered fiber probes. For this purpose tapered fiber probes were prepared and coated with a detection layer containing ion-paired fluorescent pH-transducer 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (c-HPTS). A fluorescence ratiometric approach was employed based on excitation at 415 nm and 450 nm and on the comparison of the fluorescence response at 515 nm. The suitability of tapered fiber probes for local detection of pH between 5.0 and 7.8 was demonstrated. A pH sensitivity of 0.15 pH units was achieved within the pH ranges 5.0-5.9 and 7.1-7.8, and this was improved to 0.04 pH units within the pH range 5.9-7.1. Spatial resolution of the probes was better than 20 μm and a time responsewithin 15-20 s was achieved.
Permanent Link: http://hdl.handle.net/11104/0230479