Confocal microscopy for astrocyte in vivo imaging: Recycle and reuse in microscopy

Alberto Perez-Alvarez, Alfonso Araque, Eduardo D. Martin

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

In vivo imaging is one of the ultimate and fundamental approaches for the study of the brain. Two-photon laser scanning microscopy (2PLSM) constitutes the state-of-the- art technique in current neuroscience to address questions regarding brain cell structure, development and function, blood flow regulation and metabolism. This technique evolved from laser scanning confocal microscopy (LSCM), which impacted the field with a major improvement in image resolution of live tissues in the 1980's compared to widefield microscopy. While nowadays some of the unparalleled features of 2PLSM make it the tool of choice for brain studies in vivo, such as the possibility to image deep within a tissue, LSCM can still be useful in this matter. Here we discuss the validity and limitations of LSCM and provide a guide to perform high-resolution in vivo imaging of the brain of live rodents with minimal mechanical disruption employing LSCM. We describe the surgical procedure and experimental setup that allowed us to record intracellular calcium variations in astrocytes evoked by sensory stimulation, and to monitor intact neuronal dendritic spines and astrocytic processes as well as blood vessel dynamics. Therefore, in spite of certain limitations that need to be carefully considered, LSCM constitutes a useful, convenient and affordable tool for brain studies in vivo.

Original languageEnglish (US)
JournalFrontiers in Cellular Neuroscience
Issue numberAPR
DOIs
StatePublished - Apr 7 2013
Externally publishedYes

Keywords

  • Astrocyte
  • Confocal microscopy
  • Cranial window
  • Glia
  • Imaging
  • In vivo
  • Intravital
  • Two-photon

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