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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The authors are indebted to Catherine de Vitry for the SUIV and CF0ß antibodies and to David Stern for the kind gift of iniD mutants. Financial support is acknowledged from Fondation Pierre-Gilles de Gennes pour la recherche and Agence Nationale pour la Recherche ANR-08-BIOE-002 ALGOMICS.
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Johnson, X., Vandystadt, G., Bujaldon, S. et al. A new setup for in vivo fluorescence imaging of photosynthetic activity. Photosynth Res 102, 85 (2009) doi:10.1007/s11120-009-9487-2
- Chlorophyll fluorescence
- Imaging techniques
- Screening of algal colonies