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

16 Scopus citations


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
Issue number2
Publication statusPublished - 2010


All Science Journal Classification (ASJC) codes

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

Cite this