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Published on: September 5, 2013
Two-dimensional fluorescence spectroscopy - a new tool for the determination of plant cell viability
R Vaňková1, G Kuncová2, J Opatrná1
1Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 135, 165 02 Prague 6, Czech Republic e-mail: vankova@ueb.cas.cz Fax: +420-2-20390446, , , , , , CZ.
Abstract:
Two-dimensional fluorescence spectroscopy (2D-FS) has been used as a new method for determining the viability of tobacco cells (Nicotiana tabacum L.). Both horizontal beam geometry and a vertical set-up achieved with bifurcated fibres were tested. The latter arrangement enabled us to avoid the negative effect of cell sedimentation. Incubation of a tobacco BY-2 cell suspension with dimethylsulfoxide (DMSO) (0-10% v/v) resulted in cell samples differing in their viability - from fully viable (0-2% DMSO) to totally non-viable (8-10%DMSO). The validity of determining viability by means of measuring cell esterase activity by 2D-FS using fluorescein diacetate as a fluorogenic substrate was verified by comparison with microscopic evaluation of fluorescein fluorescence as well as with the routinely adopted trypan blue exclusion test.
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