Targeted labeling of neurons in a specific functional micro-domain of the neocortex by combining intrinsic signal and two-photon imaging.

Philip John O'Herron, Zhiming Shen, Zhongyang Lu, Adrien E. Schramm, Manuel Levy, Prakash Kara

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

In the primary visual cortex of non-rodent mammals, neurons are clustered according to their preference for stimulus features such as orientation(1-4), direction(5-7), ocular dominance(8,9) and binocular disparity(9). Orientation selectivity is the most widely studied feature and a continuous map with a quasi-periodic layout for preferred orientation is present across the entire primary visual cortex(10,11). Integrating the synaptic, cellular and network contributions that lead to stimulus selective responses in these functional maps requires the hybridization of imaging techniques that span sub-micron to millimeter spatial scales. With conventional intrinsic signal optical imaging, the overall layout of functional maps across the entire surface of the visual cortex can be determined(12). The development of in vivo two-photon microscopy using calcium sensitive dyes enables one to determine the synaptic input arriving at individual dendritic spines(13) or record activity simultaneously from hundreds of individual neuronal cell bodies(6,14). Consequently, combining intrinsic signal imaging with the sub-micron spatial resolution of two-photon microscopy offers the possibility of determining exactly which dendritic segments and cells contribute to the micro-domain of any functional map in the neocortex. Here we demonstrate a high-yield method for rapidly obtaining a cortical orientation map and targeting a specific micro-domain in this functional map for labeling neurons with fluorescent dyes in a non-rodent mammal. With the same microscope used for two-photon imaging, we first generate an orientation map using intrinsic signal optical imaging. Then we show how to target a micro-domain of interest using a micropipette loaded with dye to either label a population of neuronal cell bodies or label a single neuron such that dendrites, spines and axons are visible in vivo. Our refinements over previous methods facilitate an examination of neuronal structure-function relationships with sub-cellular resolution in the framework of neocortical functional architectures.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number70
StatePublished - Jan 1 2012
Externally publishedYes

Fingerprint

Neocortex
Visual Cortex
Photons
Labeling
Neurons
Optical Imaging
Imaging techniques
Mammals
Microscopy
Coloring Agents
Vision Disparity
ocular Dominance
Dendritic Spines
Dendrites
Dyes
Fluorescent Dyes
Dendritic Cells
Axons
Spine
Labels

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Targeted labeling of neurons in a specific functional micro-domain of the neocortex by combining intrinsic signal and two-photon imaging. / O'Herron, Philip John; Shen, Zhiming; Lu, Zhongyang; Schramm, Adrien E.; Levy, Manuel; Kara, Prakash.

In: Journal of visualized experiments : JoVE, No. 70, 01.01.2012.

Research output: Contribution to journalArticle

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