A new setup for in vivo fluorescence imaging of photosynthetic activity |
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Authors: | Xenie Johnson Guillaume Vandystadt Sandrine Bujaldon Francis-André Wollman Rémi Dubois Pierre Roussel Jean Alric Daniel Béal |
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Institution: | 1.Institut de Biologie Physico-Chimique,UMR 7141, CNRS et Université Pierre et Marie Curie (Paris VI),Paris,France;2.Ecole Supérieure de Physique et de Chimie Industrielles de la ville de Paris (ESPCI-Paristech),Laboratoire d’Electronique (UMR CNRS 7084),Paris Cedex 05,France;3.Fondation Pierre-Gilles de Gennes pour la Recherche,Paris,France |
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Abstract: | Here, we describe a new imaging setup able to assess in vivo photosynthetic activity. The system specifically measures time-resolved
chlorophyll fluorescence in response to light. It is composed of a fast digital camera equipped with a wide-angle lens for
the analysis of samples up to 10 × 10 cm, i.e. entire plants or petri dishes. In the choice of CCD, we have opted for a 12-bits
high frame rate 150 fps (frames per second)] at the expense of definition (640 × 480 pixels). Although the choice of digital
camera is always a compromise between these two related features, we have designed a flexible system allowing the fast sampling
of images (down to 100 μs) with a maximum spatial resolution. This image readout system, synchronized with actinic light and
saturating pulses, allows a precise determination of F
0 and F
M, which is required to monitor PSII activity. This new imaging system, together with image processing techniques, is useful
to investigate the heterogeneity of photosynthetic activity within leaves or to screen large numbers of unicellular algal
mutant colonies to identify those with subtle changes in photosynthetic electron flow. |
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