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 Bissardon¹, Solveig Reymond¹, Murielle Salomé², Lionel André², Sam Bayat¹, Peter Cloetens², 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 Bissardon¹, Marie-Pierre Isaure², Emmanuel Lesuisse³, 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.