Analysis of Fluorescent Microscopy Images to Better Understand Keratin’s Effect on Protein Recycling Pathways in Human Embryonic Kidney Cells
- WCU Author/Contributor (non-WCU co-authors, if there are any, appear on document)
- Bailey Trumm (Creator)
- Institution
- Western Carolina University (WCU )
- Web Site: http://library.wcu.edu/
- Advisor
- Heather Coan
Abstract: Protein recycling pathways and autophagy play an important role in maintaining cellular homeostasis, but these processes are also essential for cell survival during times of stress. Protein recycling pathways, such as the ubiquitin-proteasome system (UPS), are important as old, damaged, or non-functional proteins are degraded, and the resulting free amino acids can be used to synthesize new, functional proteins. The process of autophagy works to ensure cellular homeostasis by degrading old or damaged macromolecules into their monomer parts so that they can be recycled to make new materials. Fluorescent microscopy can be used to track these pathways in a lab to inform our understanding of conditions that promote cell recycling in times of stress. However, the Coan laboratory has had difficulty standardizing image analysis using fluorescent microscopy. Counting of fluorescent image data to obtain quantitative comparisons for our lab’s cell recycling and biomaterial studies has demonstrated that manual counting is subjective and prone to error. Therefore, this study aimed to investigate the usability and reliability of images obtained via fluorescence microscopy techniques that are commonly used to investigate these pathways. In this study, I compiled data from past experiments for reanalysis with a goal of determining 1) whether these data can be replicated when reanalyzed (manually counted) and 2) whether we can glean new information from the dataset through manual counts of a parameter not previously analyzed. The results suggest that manual counting of fluorescent images can be standardized, but great care must be taken to ensure that high quality images are taken during experiments and that each manual count must follow a defined protocol. I also found that a new analysis of data may yield important results previously overlooked in our laboratory.
Analysis of Fluorescent Microscopy Images to Better Understand Keratin’s Effect on Protein Recycling Pathways in Human Embryonic Kidney Cells
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Created on 5/1/2024
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Additional Information
- Publication
- Other
- Language: English
- Date: 2024
- Keywords
- protein recycling pathways, autophagy, fluorescent microscopy