ELECTRONIC SPECTROSCOPY

The Service de Bioénergétique, Biologie Structurale et Mécanismes(SB2SM) / CNRS-UMR9198, located at JOLIOT/I2BC/CEA-Saclay, has a platform dedicated to the electronic spectroscopy of biological samples.  Historically, we specialised in the study of photosynthesis/photophysics and over the years have accumulated a diverse range of commercially available spectrometers as well as home-built optical set-ups including a “Machine de Joliot”. In general, we are able to monitor spectroscopic changes at the protein level in intact cells on many time scales for research projects that wished to monitor absorption/fluorescence changes.

A special set-up^1-3 was developed to monitor small flash-induced transient absorption changes with a time resolution of 300 ps in single flash experiments, or with a very modest number of averaged experiments.

Commercial spectrometers are available to measure chlorophyll fluorescence, P700 oxidation and NADPH fluorescence. Chlorophyll fluorescence and P700 can be measuredin vivousing leaves or algae/cyanobacteria cultures. NADPH fluorescence can be measured in algae/cyanobacteria cultures or in isolated intact chloroplasts.

Thermoluminescence can be measured with a home-built set-up that is able to monitor charge recombination reactions in photosystem II. This method can be used for leaves, algae/cyanobacteria cultures as well as isolated photosytem II.

The platform consists of:

• A home-built set-up for monitoring absorption changes with 300 ps time resolution at 266, 376, 403, 408, 448, 457, 473, 488, 515, 562, 639, 660-695 nm (tuneable) and 820 nm upon excitation at 355 or 532 nm
• Several home-built set-ups for monitoring small transient absorption changes on time scales from 5 ns to seconds at many different wavelengths between 260 and 1000 nm upon excitation anywhere between 200 and 2000 nm.
• Joliot spectrometer
• Dual-PAM (Chlorophyll fluorescence, P700 measurements, NADPH fluorescence), PAM-103-101 (Chlorophyll fluorescence), Multicolor PAM (Chlorophyll fluorescence)
• Thermoluminescence (Charge separation in photosystem II)
• Varian Cary 5000 absorption spectrometer (190-3200 nm), with Helium/nitrogen cryostats and pressure cell.
• Hitachi F-4500 Fluorescence spectrometer.

Bibliography

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