INSERM UA7, Synchrotron Radiation for Biomedicine (STROBE)
2 ARTICLES PUBLISHED IN JoVE
Biochemistry
Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis
Caroline Bissardon 1, Solveig Reymond 1, Murielle Salomé 2, Lionel André 2, Sam Bayat 1, Peter Cloetens 2, Sylvain Bohic 1,2
1Inserm, UA7, Synchrotron Radiation for Biomedicine (STROBE), Université Grenoble Alpes, 2ID16A Beamline, ESRF, The European Synchrotron
Presented here is a protocol for cell culture on silicon nitride membranes and plunge-freezing prior to X-ray fluorescence imaging with a synchrotron cryogenic X-ray nanoprobe. When only room temperature nano-analysis is provided, the frozen samples can be further freeze-dried. These are critical steps to obtain information on the intracellular elemental composition.
Chemistry
Biological Samples Preparation for Speciation at Cryogenic Temperature using High-Resolution X-Ray Absorption Spectroscopy
Caroline Bissardon 1, Marie-Pierre Isaure 2, Emmanuel Lesuisse 3, Mauro Rovezzi 4,5, Eric Lahera 4,5, Olivier Proux 4,5, Sylvain Bohic 1,6
1University Grenoble Alpes, INSERM UA7, Synchrotron Radiation for Biomedicine (STROBE), 2CNRS, UMR 5254, Université Pau & Pays Adour, E2S UPPA, Institut des Sciences Analytiques et de Physicochimie pour l'Environnement et les Matériaux, 3Institut Jacques Monod, UMR 7592 CNRS, Université Paris Diderot, 4OSUG, UMS 832 CNRS, Université Grenoble Alpes, 5FAME and FAME-UHD beamlines, ESRF, the European Synchrotron, 6ID16A beamline, ESRF, the European Synchrotron
This protocol presents a detailed procedure to prepare biological cryosamples for synchrotron-based X-ray absorption spectroscopy experiments. We describe all the steps required to optimize sample preparation and cryopreservation with examples of the protocol with cancer and phytoplankton cells. This method provides a universal standard of sample cryo-preparation.
