In vivo reactive neural plasticity investigation by means of correlative two photon: electron microscopy

View at Publisher| Export | Download | Add to List | More... Progress in Biomedical Optics and Imaging - Proceedings of SPIE Volume 8588, 2013, Article number 85881A Multiphoton Microscopy in the Biomedical Sciences XIII; San Francisco, CA; United States; 3 February 2013 through 5 February 2013; Code 97277 In vivo reactive neural plasticity investigation by means of correlative two photon: Electron microscopy (Conference Paper) Allegra Mascaro, A.L.a, Cesare, P.bc, Sacconi, L.d, Grasselli, G.b, Mandolesi, G.b, Maco, B.e, Knott, G.e, Huang, L.f, De Paola, V.f, Strata, P.bc, Pavone, F.S.a a European Laboratory for Non-Linear Spectroscopy, University of Florence, Sesto Fiorentino (FI), Italy b EBRI-Santa Lucia Foundation (IRCCS), Rome, Italy c Deptartment of Neuroscience and National Institute of Neuroscience, University of Turin, Turin, Italy View additional affiliations View references (30) Abstract In the adult nervous system, different populations of neurons correspond to different regenerative behavior. Although previous works showed that olivocerebellar fibers are capable of axonal regeneration in a suitable environment as a response to injury1, we have hitherto no details about the real dynamics of fiber regeneration. We set up a model of singularly axotomized climbing fibers (CF) to investigate their reparative properties in the adult central nervous system (CNS) in vivo. Time lapse two-photon imaging has been combined to laser nanosurgery2, 3 to define a temporal pattern of the degenerative event and to follow the structural rearrangement after injury. To characterize the damage and to elucidate the possible formation of new synaptic contacts on the sprouted branches of the lesioned CF, we combined two-photon in vivo imaging with block face scanning electron microscopy (FIB-SEM). Here we describe the approach followed to characterize the reactive plasticity after injury.