Acridine orange leukocyte fluorography in mice

Judd M Cahoon, Paul R Olson, Spencer Nielson, Tadashi R Miya, Peter Bankhead, J Graham McGeown, Timothy M Curtis, Balamurali K Ambati

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)15-9
Number of pages5
JournalExperimental Eye Research
Publication statusPublished - Mar 2014


  • Acridine Orange
  • Animals
  • Blood Flow Velocity
  • Cell Movement/physiology
  • Diabetes Mellitus, Type 1/physiopathology
  • Fluorescein Angiography
  • Fluorescent Dyes
  • Leukocytes/physiology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred NOD
  • Microscopy, Confocal
  • Ophthalmoscopes
  • Regional Blood Flow/physiology
  • Retinal Artery/physiology
  • Retinal Vasculitis/chemically induced
  • Retinal Vein/physiology
  • Tomography, Optical Coherence
  • Vascular Endothelial Growth Factor A/pharmacology
  • Video Recording


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