J. Chang, S. Rajaraman, L. Tetard
University of Central Florida,
Summary:NIH3T3 is a widely used fibroblast cell line that is commonly used as a model system for life science research. Most common bioassays used to track the behavior of NIH3T3 use fluorescence tags. However, most traditional immunocytological stains and biochemical assays are known to be an invasive tactic in imaging a cell. The evaluation of NIH3T3, from their healthy to apoptotic cells using techniques that avoid fluorescence tagging is lacking. Here we show that Raman confocal spectroscopy and atomic force microscopy (AFM) can be used to evaluate the chemical and mechanical properties of NIH3T3 without prior staining. To evade invasive techniques, Raman spectroscopy will be used to identify Raman peak shifts or other changes in the cell’s fingerprint. The variations in Raman bands will be exploited to create a three-dimensional render of the NIH3T3 cell line. We will complement our study with detailed mapping of the structure and mechanical properties of the cells using AFM. AFM force measurements have been shown to provide invaluable information on cellular systems. Apoptotic and healthy cells within the NIH3T3 cell line will be compared with our approach. Overall, investigating the NIH3T3 cell line will give an enhanced understanding of the biochemical activities within healthy and apoptotic cells within the NIH3T3 cell line, which will be important to monitor the evolution of the cells subjected to various environments in future studies.