title: Two-photon microscopy of dermal innervation in a human re-innervated model of skin.
abstract: When skin is injured, innervation can be severely disrupted. The subsequent re-innervation processes are poorly understood notably because of the inability to image the full meandering course of nerves with their ramifications and endings from histological slices. In this letter, we report on two-photon excitation fluorescence (TPEF) microscopy of entire human skin explants re-innervated by rodent sensory neurons labelled with the styryl dye FM1-43. TPEF imaging of nerve fibres to a depth up to roughly 300 μm within the dermis was demonstrated, allowing three-dimensional reconstruction of the neural tree structure. Endogenous second-harmonic imaging of type I fibrillar collagen was performed in parallel to TPEF imaging using the same nonlinear microscope, revealing the path of the nerves through the dermis.
authors: David
authors: D
authors: Sevrain
authors: Yann
authors: Y
authors: Le Grand
authors: Virginie
authors: V
authors: Buhé
authors: Christine
authors: C
authors: Jeanmaire
authors: Gilles
authors: G
authors: Pauly
authors: Jean-Luc
authors: JL
authors: Carré
authors: Laurent
authors: L
authors: Misery
authors: Nicolas
authors: N
authors: Lebonvallet
mesh_terms: D000818:Animals; D018920:Coculture Techniques; D020405:Dermis / Q000294:innervation*; D005456:Fluorescent Dyes; D006801:Humans; D021621:Imaging, Three-Dimensional; D036641:Microscopy, Fluorescence, Multiphoton; D008959:Models, Neurological; D009416:Nerve Regeneration; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D051381:Rats; D017208:Rats, Wistar; D011984:Sensory Receptor Cells / Q000502:physiology; D012867:Skin / Q000293:injuries* / Q000294:innervation*
chemical_list: C073804:FM1 43; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds