Article Application of multispectral imaging detects areas with neuronal myelin loss, without tissue labelling Eleftheria Vazgiouraki 1,2, † , Vassilis M. Papadakis 2,3, †, *, Paschalis Efstathopoulos 1 , Iakovos Lazaridis 1 , Ioannis Charalampopoulos 1 , Costas Fotakis 2,4 , and Achille Gravanis 1,5 1 Department of Pharmacology, School of Medicine, University of Crete, Voutes Campus, Heraklion, Crete GR-71003, Greece, 2 Institute of Electronic Structure and LASER (I.E.S.L.), Foundation for Research and Technology, Hellas (FO.R.T.H.), Nikolaou Plastira 100, Vassilika Vouton GR-70013 Heraklion, Crete, Greece, 3 Aerospace Non-Destructive Testing Laboratory, Delft University of Technology, Kluyverweg 1 (building 62) 2629 HS Delft, The Netherlands, 4 Department of Physics, School of Science and Engineer- ing, University of Crete, Voutes Campus, Heraklion, Crete GR-71003, Greece, and 5 Institute of Molecular Biology and Biotechnology (I.M.B.B.), Foundation for Research and Technology, Hellas (FO.R.T.H.), Nikolaou Plastira 100, Vassilika Vouton GR-70013 Heraklion, Crete, Greece *To whom correspondence should be addressed. E-mail: billyp@iesl.forth.gr † These authors contributed equally to this work. Received 19 March 2015; Accepted 5 October 2015 Abstract The application of multispectral imaging to discriminate myelinated and demyelinated areas of neural tissue is herein presented. The method is applied through a custom-made, multispectral imaging monochromator, coupled to a commercially available microscope. In the present work, a series of spinal cord sections were analysed derived from mice with experimental autoimmune encephalomyelitis (EAE), an experimental model widely used to study multiple sclerosis (MS). The multispectral microscope allows imaging of local areas with loss of myelin without the need of tissue labelling. Imaging with the aforementioned method and system is compared in a parallel way with conventional methods (wide-field and confocal fluorescence microscopies). The diagnostic sensitivity of our method is 90.4% relative to the ‘gold standard’ method of immunofluorescence microscopy. The presented method offers a new platform for the possible future development of an in vivo, real-time, non-invasive, rapid imaging diagnostic tool of spinal cord myelin loss-derived pathologies. Key words: imaging, microscopy, EAE, wide-field microscope, confocal microscope, neuronal degeneration Introduction Multiple sclerosis (MS) is characterized by inflammation and demyelinating lesions in white matter (WM) of the central nervous system (CNS) [1], leading to permanent neurological disabilities (sensory and motor deficiencies) [2,3]. Research on MS has been performed mainly with the Microscopy, 2015, 1–10 doi: 10.1093/jmicro/dfv349 © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com 1 Microscopy Advance Access published October 28, 2015 at University of Crete on November 6, 2015 http://jmicro.oxfordjournals.org/ Downloaded from