Intracellular and Juxtacellular Staining with Biocytin

Charles J. Wilson1, R.N.S. Sachdev1

1 The University of Texas at San Antonio, San Antonio, Texas
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 1.12
DOI:  10.1002/0471142301.ns0112s26
Online Posting Date:  May, 2004
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Many physiological studies require microscopic examination of the recorded neuron for identification. This unit describes how intracellular and extracellular recording can be combined with single‐neuron staining to enable sequential physiological and morphological studies.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Basic Protocol 1: Intracellular Biocytin Staining with Sharp Electrodes
  • Alternate Protocol 1: Intracellular Biocytin Staining with Patch Electrodes
  • Basic Protocol 2: Juxtacellular Biocytin Staining
  • Support Protocol 1: Tissue Processing
  • Commentary
  • Literature Cited
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Intracellular Biocytin Staining with Sharp Electrodes

  Materials
  • Tissues
  • 0.1% Triton‐X 100
  • Phosphate‐buffered saline (PBS), pH 7.2
  • ABC kit or avidin‐conjugated fluorescent marker (e.g., avidin‐Texas Red)
  • 0.05% (v/v) 3′,3′‐diaminobenzidine (DAB)
  • 3% (v/v) hydrogen peroxide
  • Vibratome or freezing microtome
  • Shaker
  • Additional reagents and equipment for dehydrating and mounting sections (unit 1.1)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Bevan, M.D., Booth, P.A., Eaton, S.A., and Bolam, J.P. 1998. Selective innervation of neostriatal interneurons by a subclass of neuron in the globus pallidus of the rat. J. Neurosci. 18:9438‐9452.
   Horikawa, K. and Armstrong W. 1988. A versatile means of intracellular labeling: Injection of biocytin and its detection with avidin conjugates. J. Neurosci. Methods 25:1‐11.
   Kawaguchi, Y., Wilson, C.J., and Emson, P.C. 1989. Intracellular recording of identified neostriatal patch and matrix spiny cells in a slice preparation preserving cortical inputs. J. Neurophysiol. 62:1052‐1068.
   Kawaguchi, Y. 1992. Large aspiny cells in the matrix of the rat neostriatum in vitro: Physiological identification, relation to the compartments and excitatory postsynaptic currents. J. Neurophysiol. 67:1669‐1682.
   Kawaguchi, Y. 1993. Physiological, morphological, and histochemical characterization of three classes of interneurons in rat neostriatum. J. Neurosci. 13:4908‐4923.
   Nisenbaum, E.S. and Wilson, C.J. 1995. Potassium currents responsible for inward and outward rectification in rat neostriatal spiny projection neurons. J. Neurosci. 15:4449‐4463.
   Pinault, D. 1994. Golgi‐like labeling of a single neuron recorded extracellularly. Neurosci. Lett. 11:255‐260
   Pinault, D. 1996. A novel single‐cell staining procedure performed in vivo under electrophysiological control: Morpho‐functional features of juxtacellularly labeled thalamic cells and other central neurons with biocytin or Neurobiotin. J. Neurosci. Methods 65:113‐136.
   Xu, Z.C., Wilson, C.J., and Emson, P.C. 1992. Morphology of intracellularly stained spiny neurons in rat striatal grafts. Neuroscience 48:95‐110.
   Zheng, T. and Wilson, C.J. 2002. Corticostriatal combinatorics: The implications of corticostriatal axonal arborizations. J. Neurophysiol. 87:1007‐1017.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library