Photo- and bio-physical characterization of novel violet and near-infrared lipophilic fluorophores for neuronal tracing

J. Tonniges, M. Hansen, J. Duncan, M. J. Bassett, B. Fritzsch, B. D. Gray, A. Easwaran, Michael G. Nichols

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)117-134
Number of pages18
JournalJournal of Microscopy
Volume239
Issue number2
DOIs
Publication statusPublished - 2010

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All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology
  • Medicine(all)

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