Fluorescence time-lapse imaging of single cells targeted with a focused scanning charged-particle microbeam

S. Bourret (1) , F. Vianna (1, 2) , Guillaume Devès (1) , V. Atallah , P. Moretto (1) , H. Seznec (1) , P. Barberet (1)

1 IPCV - Interface Physique et Chimie pour le Vivant
2 IRSN - Institut de Radioprotection et de Sûreté Nucléaire

Charged particle microbeams provide unique features to study targeted and non-targeted radiation response and have recently emerged as a powerful tool to investigate radiation-induced DNA damage and repair. We have developed a charged particle microbeam delivering protons and alpha particles in the MeV energy range equipped with online time-lapse imaging capabilities. The beam is focused to a sub-micrometer beam spot under vacuum by means of a triplet of magnetic quadrupoles and extracted in air through a 200 nm Si3N4 window. The end-station is equipped with an automated fluorescence microscope used for single cell targeting and online time-lapse imaging. Cells are kept in their medium during the irradiation procedure and the sample temperature is regulated to 37 C. An overall targeting accuracy of 2.0 ± 0.7 lm has been measured by tracking the re-localization of the XRCC1 protein. First measurements of this re-localization shows the ability of our system to follow online the radiationinduced re-localization of proteins in the first minutes after irradiation.

Mots clés: Microbeam Single cell Time-lapse microscopy Targeted irradiation Radiation biology

Domaines: Instrumentations et Détecteurs [physics.ins-det]

S. Bourret, F. Vianna, Guillaume Devès, V. Atallah, P. Moretto, et al.. Fluorescence time-lapse imaging of single cells targeted with a focused scanning charged-particle microbeam. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, 2014, 325, pp.27-34. ⟨10.1016/j.nimb.2014.02.004⟩. ⟨in2p3-00956455⟩