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.

Institute of Chemical Process Fundamentals Academy of Sciences of the Czech Republic Rozvojová 135 165 02 Prague 6 Czech Republic

Institute of Experimental Botany Academy of Sciences of the Czech Republic Rozvojová 135 165 02 Prague 6 Czech Republic e mail Fax 420 2 20390446

Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic Flemingovo nám 2 166 10 Prague 6 Czech Republic

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