In vivo local dye electroporation for Ca2+ imaging and neuronal-circuit tracing

Shin Nagayama, Max Fletcher, Wenhui Xiong, Xiaohua Lu, Shaoqun Zeng, Wei R. Chen

Research output: Contribution to journalArticle

Abstract

A major technical challenge for using optical imaging to analyze neuronal circuit functions is how to effectively load synthetic Ca2+ dyes or neural tracers into the brain. We introduce here a simple but versatile approach to label many neurons and clearly visualize their axonal and dendritic morphology. The method uses a large-tip patch pipette filled with dextran-conjugated Ca2+ dyes or fluorescent tracers. By inserting the pipette into a targeted brain area and passing microampere current pulses, dyes or tracers are electroporated into dendrites and axons near the pipette tip. The dyes are then transported to reveal the entire cell morphology, suitable for both functional Ca2+ imaging and neuronal circuit tracing. This process requires basic physiological equipment normally available in a physiological laboratory.

Original languageEnglish (US)
Pages (from-to)940-947
Number of pages8
JournalCold Spring Harbor Protocols
Volume2014
Issue number9
DOIs
StatePublished - Jan 1 2014

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Electroporation
Coloring Agents
Imaging techniques
Networks (circuits)
Brain
Optical Imaging
Dendrites
Dextrans
Fluorescent Dyes
Axons
Neurons
Labels
Equipment and Supplies

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

In vivo local dye electroporation for Ca2+ imaging and neuronal-circuit tracing. / Nagayama, Shin; Fletcher, Max; Xiong, Wenhui; Lu, Xiaohua; Zeng, Shaoqun; Chen, Wei R.

In: Cold Spring Harbor Protocols, Vol. 2014, No. 9, 01.01.2014, p. 940-947.

Research output: Contribution to journalArticle

Nagayama, Shin ; Fletcher, Max ; Xiong, Wenhui ; Lu, Xiaohua ; Zeng, Shaoqun ; Chen, Wei R. / In vivo local dye electroporation for Ca2+ imaging and neuronal-circuit tracing. In: Cold Spring Harbor Protocols. 2014 ; Vol. 2014, No. 9. pp. 940-947.
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