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

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Kasík, Ivan Podrazký, Ondrej Mrázek, Jan Martan, Tomás Matejec, Vlastimil Hoyerová, Klára Kamínek, Miroslav
Description	Author Affiliation: Kasík I ( Institute of Photonics and Electronics, AS CR, v.v.i., Chaberska 57, Prague 8 182 51, Czech Republic. Electronic address: kasik@ufe.cz.)
Abstract	Minimally invasive in vivo measurement of pH in microscopic biological samples of µm or 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 pH5.0 to pH7.8 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 response within 15-20s was achieved. Despite the minute dimensions of the tapered fiber probes the setup developed was relatively robust and compact in construction and performed reliably. It has been successfully employed for the in vivo local determination of pH of mechanically resistant plant tissues of A. thaliana of microscopic scale. The detection of momentary pH gradients across the intact plant seems to be a good tool for the determination of changes in pH in response to experimental treatments affecting for example enzyme activities, availability of mineral nutrients, hormonal control of plant development and plant responses to environmental cues.
File Format	HTM / HTML
ISSN	09284931
Issue Number	8
Journal	Materials Science and Engineering: C
Volume Number	33
e-ISSN	18730191
Language	English
Publisher	Elsevier
Publisher Date	2013-12-01
Publisher Place	Netherlands
Access Restriction	Subscribed
Subject Keyword	Discipline Materials Science Arabidopsis Metabolism Arylsulfonates Chemistry Fluorescent Dyes Hydrogen-ion Concentration Spectrometry, Fluorescence Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Mechanics of Materials Biomaterials Condensed Matter Physics Bioengineering Mechanical Engineering

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