22 February 2013 In vivo reactive neural plasticity investigation by means of correlative two photon: electron microscopy
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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.
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A. L. Allegra Mascaro, A. L. Allegra Mascaro, P. Cesare, P. Cesare, L. Sacconi, L. Sacconi, G. Grasselli, G. Grasselli, G. Mandolesi, G. Mandolesi, B. Maco, B. Maco, G. Knott, G. Knott, L. Huang, L. Huang, V. De Paola, V. De Paola, P. Strata, P. Strata, F. S. Pavone, F. S. Pavone, } "In vivo reactive neural plasticity investigation by means of correlative two photon: electron microscopy", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85881A (22 February 2013); doi: 10.1117/12.2005953; https://doi.org/10.1117/12.2005953
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