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Combining microiontophoresis and multiphoton excitation microscopy to investigate neuronal signaling

Hartveit, Espen; Veruki, Margaret Lin
Chapter, Peer reviewed
Accepted version
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URI
https://hdl.handle.net/1956/22833
Date
2019
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  • Department of Biomedicine [447]
Original version
https://doi.org/10.1007/978-1-4939-9702-2_7
Abstract
Multiphoton excitation (MPE) microscopy allows subcellular structural and functional imaging of neurons and can be combined with techniques for activating postsynaptic receptors at spatial and temporal scales that mimic normal synaptic transmission. Here, we describe procedures for combining MPE imaging of dye-filled neurons with fast microiontophoresis, by which neurotransmitter agonists can be applied from high-resistance micropipettes with subcellular resolution. With adequate compensation of the pipette capacitance, the effective time constant of the pipette is reduced, and this permits application of very brief pulses of receptor agonist (≤1 ms). The consequent high temporal and spatial resolution leads to the high specificity required for single-synapse investigations. This chapter includes detailed procedures for electrophysiological whole-cell recording, structural and functional (Ca2+) MPE imaging of dye-filled neurons, targeting a microiontophoresis pipette to a specific subcellular compartment of a dye-filled neuron under visual control, and capacitance compensation of the microiontophoresis pipette, as well as examples of experimental results that can be obtained.
Description
Under embargo until: 2021-11-01
Publisher
Springer
Series
Neuromethods
Copyright
Copyright Springer Science+Business Media, LLC, part of Springer Nature 2019

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