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Exp Eye Res. 2014 Mar;120:15-9. doi: 10.1016/j.exer.2013.12.002. Epub 2013 Dec 12.

Acridine orange leukocyte fluorography in mice.

Author information

  • 1Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States. Electronic address: judd.cahoon@hsc.utah.edu.
  • 2Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States.
  • 3Centre for Vision and Vascular Science, Queen's University of Belfast, Institute of Clinical Sciences, Block A, Royal Victoria Hospital Belfast, Ireland.

Abstract

Simultaneous non-invasive visualization of blood vessels and nerves in patients can be obtained in the eye. The retinal vasculature is a target of many retinopathies. Inflammation, readily manifest by leukocyte adhesion to the endothelial lining, is a key pathophysiological mechanism of many retinopathies, making it a valuable and ubiquitous target for disease research. Leukocyte fluorography has been extensively used in the past twenty years; however, fluorescent markers, visualization techniques, and recording methods have differed between studies. The lack of detailed protocol papers regarding leukocyte fluorography, coupled with lack of uniformity between studies, has led to a paucity of standards for leukocyte transit (velocity, adherence, extravasation) in the retina. Here, we give a detailed description of a convenient method using acridine orange (AO) and a commercially available scanning laser ophthalmoscope (SLO, HRA-OCT Spectralis) to view leukocyte behavior in the mouse retina. Normal mice are compared to mice with acute and chronic inflammation. This method can be readily adopted in many research labs.

Copyright © 2013 Elsevier Ltd. All rights reserved.

KEYWORDS:

acridine orange; blood flow; inflammation; leukocyte; retina

PMID:
24333760
[PubMed - indexed for MEDLINE]
PMCID:
PMC3943592
Free PMC Article
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