Stable in vivo imaging of densely populated glia, axons and blood vessels in the mouse spinal cord using two-photon microscopy

J Neurosci Methods. 2008 Mar 30;169(1):1-7. doi: 10.1016/j.jneumeth.2007.11.011. Epub 2007 Nov 28.

Abstract

In vivo imaging has revolutionized our understanding of biological processes in brain physiology and pathology. However, breathing-induced movement artifacts have impeded the application of this powerful tool in studies of the living spinal cord. Here we describe in detail a method to image stably and repetitively, using two-photon microscopy, the living spinal tissue in mice with dense fluorescent cells or axons, without the need for animal intubation or image post-processing. This simplified technique can greatly expand the application of in vivo imaging to study spinal cord injury, regeneration, physiology and disease.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acepromazine / pharmacology
  • Anesthesia / methods*
  • Anesthetics / pharmacology
  • Animals
  • Axons / physiology
  • Axons / ultrastructure
  • Capillaries / cytology
  • Capillaries / physiology
  • Cell Movement / physiology
  • Green Fluorescent Proteins / genetics*
  • Image Processing, Computer-Assisted / methods*
  • Ketamine / pharmacology
  • Laminectomy / methods
  • Mice
  • Mice, Transgenic
  • Microcirculation / cytology
  • Microcirculation / physiology
  • Microglia / cytology
  • Microglia / physiology
  • Microscopy, Confocal / methods*
  • Neurosurgical Procedures / methods
  • Respiratory Physiological Phenomena / drug effects
  • Spinal Cord / blood supply
  • Spinal Cord / cytology*
  • Staining and Labeling / methods*
  • Stereotaxic Techniques / instrumentation
  • Urethane / pharmacology
  • Xylazine / pharmacology

Substances

  • Anesthetics
  • Green Fluorescent Proteins
  • Xylazine
  • Urethane
  • Acepromazine
  • Ketamine