Practical methods for in vivo cortical physiology with 2-photon microscopy and bulk loading of fluorescent calcium indicator dyes

Stephen D.Van Hooser, Elizabeth N. Johnson, Ye Li, Mark Mazurek, Julie H. Culp, Arani Roy, Rishabh Kasliwal, Kelly Flavahan

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

In vivo 2-photon imaging of neurons that have been bulk-loaded with fl uorescent calcium indicator dyes is permitting many fundamental principles of neural circuit organization and development to be uncovered. In this article, we describe the materials and procedures that we have used in our investigations of ferrets, tree shrews, and mice. Special attention is given to the design and construction of custom stereotaxic devices and the prevention of stray light from entering the 2-photon microscope during vision experiments.

Original languageEnglish (US)
Title of host publicationNeural Tracing Methods
Subtitle of host publicationTracing Neurons and Their Connections
PublisherSpringer New York
Pages117-141
Number of pages25
Volume92
ISBN (Electronic)9781493919635
ISBN (Print)9781493919628
DOIs
StatePublished - Jan 1 2015

Publication series

NameNeuromethods
PublisherHumana Press
ISSN (Print)0893-2336

    Fingerprint

Keywords

  • 2-photon
  • AM calcium dyes
  • AM dyes
  • Epifluorescence
  • Head plate
  • Headplate
  • Light block
  • Optical chamber
  • Oregon Green BAPTA-1
  • Two-photon

Cite this

Hooser, S. D. V., Johnson, E. N., Li, Y., Mazurek, M., Culp, J. H., Roy, A., Kasliwal, R., & Flavahan, K. (2015). Practical methods for in vivo cortical physiology with 2-photon microscopy and bulk loading of fluorescent calcium indicator dyes. In Neural Tracing Methods: Tracing Neurons and Their Connections (Vol. 92, pp. 117-141). (Neuromethods). Springer New York. https://doi.org/10.1007/978-1-4939-1963-5_6