Formal UROP Report

Abstract

Focal adhesions are structures that secure the cell to the surrounding extracellular matrix. Immunofluorescence staining of vinculin is a common technique that allows for quantification of focal adhesion size and quantity. A protocol for staining focal adhesions in vascular smooth muscle cells (VSMCs) was synthesized through a review of relevant literature and optimized for performance. Process optimization demonstrated that an incubation period in 2% bovine serum albumin prevents non-specific binding, therefore, allowing for the mouse vinculin monoclonal antibody to bind more readily to vinculin and vinculin only in VSMCs. ImageJ software was used to measure an average focal adhesion length of 32.302 𝜇m in standard VSMCs. The optimized protocol for immunofluorescence labeling of vinculin will be used in future applications and by other lab personnel to better inform our understanding of cellular structures that contribute to mechanical integrity.