title: Photo- and bio-physical characterization of novel violet and near-infrared lipophilic fluorophores for neuronal tracing.
abstract: Lipophilic fluorescent dyes have been used to trace neuronal connections because of their ability to diffuse laterally within nerve cell membranes. Given the hundreds to thousands of connections that a typical neuron makes with its neighbours, a diffusion-matched set of spectrally distinct dyes is desirable. To extend a set of these dyes to obtain six independent labels, we have characterized the properties of novel violet and near-infrared candidates. By combining two-photon and confocal microscopy all of these candidates can be imaged using a single Titanium Sapphire laser. Here we present measurements of the two-photon action cross-sections and diffusion properties of the dyes, using either the relative diffusion distance or fluorescence recovery after photobleaching techniques, and demonstrate six-colour neuronal tracing within the spinal cord and brain tissue.
authors.affiliation: Creighton University, Department of Physics, Omaha, NE 68178, USA.
authors.forename: J
authors.initials: J
authors.lastname: Tonniges
authors.forename: M
authors.initials: M
authors.lastname: Hansen
authors.forename: J
authors.initials: J
authors.lastname: Duncan
authors.forename: M J
authors.initials: MJ
authors.lastname: Bassett
authors.forename: B
authors.initials: B
authors.lastname: Fritzsch
authors.forename: B D
authors.initials: BD
authors.lastname: Gray
authors.forename: A
authors.initials: A
authors.lastname: Easwaran
authors.forename: M G
authors.initials: MG
authors.lastname: Nichols
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology; D005456:Fluorescent Dyes / Q000494:pharmacology*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D009474:Neurons / Q000166:cytology*; D013116:Spinal Cord / Q000166:cytology; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes