Isolation of Mouse Retinal Capillaries and Subendothelial Matrix for Stiffness Measurement Using Atomic Force MicroscopyIrene Santiago Tierno 1,2,3, Mahesh Agarwal 1,2, Nikolaos Matisioudis 2,4, Sathishkumar Chandrakumar 1,2, Kaustabh Ghosh 1,2,3
1Department of Ophthalmology, University of California, Los Angeles, 2Doheny Eye Institute, 3Molecular, Cellular, and Integrative Physiology Interdepartmental PhD Program, University of California, Los Angeles, 4Department of Bioengineering, University of Pennsylvania
We recently identified retinal capillary stiffening as a new paradigm for retinal dysfunction associated with diabetes. This protocol elaborates the steps for isolation of mouse retinal capillaries and the subendothelial matrix from retinal endothelial cultures, followed by a description of the stiffness measurement technique using atomic force microscopy.