Vascular Perfusion and Retinal Leukocyte Quantification

Blood was removed from the vasculature of anesthetized animals by perfusion with phosphate-buffered saline (PBS; 10 mM phosphate buffer, 2.7 mM potassium chloride and 137 mM sodium chloride, pH 7.4; Sigma-Aldrich, St. Louis, MO) via a heart catheter. Animals were then perfused with fluorescein-coupled concanavalin A lectin (20 μg/ml in PBS; Vector Laboratories, Burlingame, CA), as described previously [20 (link),23 (link),26 (link)]. Flat-mounted retinas were imaged via fluorescence microscopy, and the number of leukocytes adherent to the vascular wall was counted.

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Saliba A., Du Y., Liu H., Patel S., Roberts R., Berkowitz B.A, & Kern T.S. (2015). Photobiomodulation Mitigates Diabetes-Induced Retinopathy by Direct and Indirect Mechanisms: Evidence from Intervention Studies in Pigmented Mice. PLoS ONE, 10(10), e0139003.

Publication 2015

concanavalin A lectin

Animals Blood Buffer Concanavalin a Fluorescein Fluorescence microscopy Heart catheter Lectin Leukocytes number Perfusion Phosphate Potassium chloride Retinas Saline Sodium chloride Vascular Vector

Corresponding Organization : Veterans Health Administration

Other organizations : Universidade Católica de Brasília, Wayne State University, Michigan United

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Variable analysis

independent variables

Perfusion with phosphate-buffered saline (PBS)
Perfusion with fluorescein-coupled concanavalin A lectin

dependent variables

Number of leukocytes adherent to the vascular wall

control variables

Anesthetized animals
Heart catheter
Flat-mounted retinas
Fluorescence microscopy

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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