Using TIRF (Total Internal Reflection Fluorescence) microscopy to measure actin filaments
Baker, Haleigh, Fater, Aaron, Beck, Moriah R.. Using TIRF (Total Internal Reflection Fluorescence) microscopy to measure actin filaments. -- Fyre in STEM Showcase, 2022.
Actin is a protein that can maintain the shape of cells and is responsible for cell motility. Actin exists as monomers that then can be polymerized into bundles or filaments. Actin has been associated with metastatic cancer because cells rely on actin polymerization for normal cell motility, and this is enhanced in this invasive type of cell motility. The relationship between actin polymerization and cell motility is that actin will form filaments that provide support and propel cell movement. Metastatic cells require fast polymerization of actin that also form an interconnected network for added strength. Previous work in the Beck lab has relied upon freshly prepared actin and want to move towards studying frozen actin to facilitate using stored preparations of actin. We hypothesize that frozen actin will polymerize the same as freshly prepared actin. We can track this polymerization visually by using TIRF (Total Internal Reflection Fluorescence) Microscopy to compare the polymerization between both fresh and frozen actin. With TIRF microscopy we take images every 5 seconds over 20 minutes and then use these images to measure and track the growth of actin over time to determine the rate of polymerization. From the data collected so far, we have determined that our frozen actin polymerizes half as fast as fresh actin. Using this knowledge, it is likely that we cannot use our frozen actin in place of fresh actin.
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Research project completed at the Department of Chemistry and Biochemistry.