Primer to Voltage Imaging With ANNINE Dyes and Two-Photon Microscopy
| Autor: | Christopher J. Roome, Bernd Kuhn |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Materials science solvatochromism Voltage-sensitive dye Review lcsh:RC321-571 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine Calcium imaging Two-photon excitation microscopy Microscopy electrochromism medicine VSD ANNINE lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry two-photon Membrane potential Electroporation voltage-sensitive dye Electrophysiology 030104 developmental biology medicine.anatomical_structure Biophysics Neuron membrane potential 030217 neurology & neurosurgery Neuroscience voltage imaging |
| Zdroj: | Frontiers in Cellular Neuroscience, Vol 13 (2019) Frontiers in Cellular Neuroscience |
| ISSN: | 1662-5102 |
| DOI: | 10.3389/fncel.2019.00321 |
| Popis: | ANNINE-6 and ANNINE-6plus are voltage-sensitive dyes that when combined with two-photon microscopy are ideal for recording of neuronal voltages in vivo, in both bulk loaded tissue and the dendrites of single neurons. Here, we describe in detail but for a broad audience the voltage sensing mechanism of fast voltage-sensitive dyes, with a focus on ANNINE dyes, and how voltage imaging can be optimized with one-photon and two-photon excitation. Under optimized imaging conditions the key strengths of ANNINE dyes are their high sensitivity (0.5%/mV), neglectable bleaching and phototoxicity, a linear response to membrane potential, and a temporal resolution which is faster than the optical imaging devices currently used in neurobiology (order of nanoseconds). ANNINE dyes in combination with two-photon microscopy allow depth-resolved voltage imaging in bulk loaded tissue to study average membrane voltage oscillations and sensory responses. Alternatively, if ANNINE-6plus is applied internally, supra and sub threshold voltage changes can be recorded from dendrites of single neurons in awake animals. Interestingly, in our experience ANNINE-6plus labeling is impressively stable in vivo, such that voltage imaging from single Purkinje neuron dendrites can be performed for 2 weeks after a single electroporation of the neuron. Finally, to maximize their potential for neuroscience studies, voltage imaging with ANNINE dyes and two-photon microscopy can be combined with electrophysiological recording, calcium imaging, and/or pharmacology, even in awake animals. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|