title: Acridine orange leukocyte fluorography in mice.
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.
authors: Department 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.
authors: Judd M
authors: JM
authors: Cahoon
authors: Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States.
authors: Paul R
authors: PR
authors: Olson
authors: Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States.
authors: Spencer
authors: S
authors: Nielson
authors: Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States.
authors: Tadashi R
authors: TR
authors: Miya
authors: Centre for Vision and Vascular Science, Queen's University of Belfast, Institute of Clinical Sciences, Block A, Royal Victoria Hospital Belfast, Ireland.
authors: Peter
authors: P
authors: Bankhead
authors: Centre for Vision and Vascular Science, Queen's University of Belfast, Institute of Clinical Sciences, Block A, Royal Victoria Hospital Belfast, Ireland.
authors: J Graham
authors: JG
authors: McGeown
authors: Centre for Vision and Vascular Science, Queen's University of Belfast, Institute of Clinical Sciences, Block A, Royal Victoria Hospital Belfast, Ireland.
authors: Timothy M
authors: TM
authors: Curtis
authors: Department of Ophthalmology and Visual Science, John A Moran Eye Center, University of Utah, Salt Lake City, UT 84109, United States.
authors: Balamurali K
authors: BK
authors: Ambati
mesh_terms: D000165:Acridine Orange; D000818:Animals; D001783:Blood Flow Velocity; D002465:Cell Movement / Q000502:physiology; D003922:Diabetes Mellitus, Type 1 / Q000503:physiopathology; D005451:Fluorescein Angiography; D005456:Fluorescent Dyes; D007962:Leukocytes / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D018613:Microscopy, Confocal; D019721:Ophthalmoscopes; D012039:Regional Blood Flow / Q000502:physiology; D012161:Retinal Artery / Q000502:physiology*; D031300:Retinal Vasculitis / Q000139:chemically induced / Q000503:physiopathology; D012169:Retinal Vein / Q000502:physiology*; D041623:Tomography, Optical Coherence; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D014741:Video Recording
chemical_list: D005456:Fluorescent Dyes; D042461:Vascular Endothelial Growth Factor A; D000165:Acridine Orange
keywords: acridine orange; blood flow; inflammation; leukocyte; retina