The pitfalls of in vivo imaging techniques: evidence for cellular damage caused by synchrotron X-ray computed micro-tomography.

Francesco Petruzzellis, Chiara Pagliarani, Tadeja Savi, Adriano Losso, Silvia Cavalletto, Giuliana Tromba, Christian Dullin, Andreas Bär, Andrea Ganthaler, Andrea Miotto, Stefan Mayr, Maciej A Zwieniecki, Andrea Nardini, Francesca Secchi

Journal: The New phytologist 2019;220(1):104-110

PMID: 30040128

Abstract

Synchrotron X-ray computed micro-tomography (microCT) has emerged as a promising noninvasive technique for in vivo monitoring of xylem function, including embolism build-up under drought and hydraulic recovery following re-irrigation. Yet, the possible harmful effects of ionizing radiation on plant tissues have never been quantified. We specifically investigated the eventual damage suffered by stem living cells of three different species exposed to repeated microCT scans. Stem samples exposed to one, two or three scans were used to measure cell membrane and RNA integrity, and compared to controls never exposed to X-rays. Samples exposed to microCT scans suffered serious alterations to cell membranes, as revealed by marked increase in relative electrolyte leakage, and also underwent severe damage to RNA integrity. The negative effects of X-rays were apparent in all species tested, but the magnitude of damage and the minimum number of scans inducing negative effects were species-specific. Our data show that multiple microCT scans lead to disruption of fundamental cellular functions and processes. Hence, microCT investigation of phenomena that depend on physiological activity of living cells may produce erroneous results and lead to incorrect conclusions.

© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

Address: Dipartimento di Scienze della Vita, Università di Trieste, Via L. Giorgieri 10, 34127, Trieste, Italy.; Institute for Sustainable Plant Protection, National Research Council, Strada delle Cacce 73, 10135, Torino, Italy.; Dipartimento di Scienze Agrarie, Forestali e Alimentari, Università di Torino, Largo Paolo Braccini 2, 10095, Grugliasco, TO, Italy.; Division of Viticulture and Pomology, Department of Crop Sciences, University of Natural Resources and Life Sciences, Konrad Lorenz Straße 24, A-3430, Tulln, Vienna, Austria.; Department of Botany, University of Innsbruck, Sternwartestraße 15, 6020, Innsbruck, Austria.; Elettra Sincrotrone Trieste, Area Science Park, 34149, Basovizza, Trieste, Italy.; Institute for Diagnostic and Interventional Radiology, University Medical Center Göttingen, Robert-Koch-Straße 40, 37075, Göttingen, Germany.; Max-Plank-Institute for Experimental Medicine, Hermann-Rein-Straße 3, 37075, Göttingen, Germany.; Department of Plant Sciences, University of California Davis, One Shields Ave, Davis, CA, 95616, USA.
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