Rose bengal photothrombosis by confocal optical imaging in vivo: A model of single vessel stroke

Lora Talley Watts, Wei Zheng, R. Justin Garling, Victoria C. Frohlich, James Donald Lechleiter

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel. The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke.

Original languageEnglish (US)
Article numbere52794
JournalJournal of Visualized Experiments
Volume2015
Issue number100
DOIs
StatePublished - Jun 23 2015

Keywords

  • In vivo microscopy
  • Issue 100
  • Lacunar stroke
  • Medicine
  • Photothrombosis
  • Rose Bengal
  • Silent stroke
  • Single vessel stroke

ASJC Scopus subject areas

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

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