Photo- and bio-physical characterization of novel violet and near-infrared lipophilic fluorophores for neuronal tracing
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 d...
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| Veröffentlicht in: | Journal of microscopy (Oxford) 2010-08, Vol.239 (2), p.117-134 |
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| container_title | Journal of microscopy (Oxford) |
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| creator | TONNIGES, J HANSEN, M DUNCAN, J BASSETT, M.J FRITZSCH, B GRAY, B.D EASWARAN, A NICHOLS, M.G |
| description | 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. |
| doi_str_mv | 10.1111/j.1365-2818.2009.03363.x |
| format | Article |
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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. 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| ispartof | Journal of microscopy (Oxford), 2010-08, Vol.239 (2), p.117-134 |
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| language | eng |
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| source | Wiley Free Content; MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Animals Brain - cytology Fluorescent Dyes - pharmacology FRAP lipophilic dyes membrane diffusion Mice Microscopy, Confocal - methods neuroanatomical tracing Neurons - cytology Spinal Cord - cytology Staining and Labeling - methods two-photon excitation |
| title | Photo- and bio-physical characterization of novel violet and near-infrared lipophilic fluorophores for neuronal tracing |
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