Year-end Sale % OFF 
Abstract
Live cell Raman micro-spectroscopy is emerging as a promising bioanalytical technique for label-free discrimination of a range of different cell types (e.g. cancer cells and fibroblasts) and behaviors (e.g. apoptosis). The aim of this study was to determine whether confocal Raman micro-spectroscopy shows sufficient sensitivity and specificity for identification of primary human bronchial epithelial cells (HBECs) to be used for live cell biological studies in vitro. We first compared cell preparation substrates and media, considering their influence on lung cell proliferation and Raman spectra, as well as methods for data acquisition, using different wavelengths (488 nm, 785 nm) and scan protocols (line, area). Evaluating these parameters using human lung cancer (A549) and fibroblast (MRC5) cell lines confirmed that line-scan data acquisition at 785 nm using complete cell media on a quartz substrate gave optimal performance. We then applied our protocol to acquisition of data from primary human bronchial epithelial cells (HBEC) derived from three independent sources, revealing an average sensitivity for different cell types of 96.3% and specificity of 95.2%. These results suggest that Raman micro-spectroscopy is suitable for delineating primary HBEC cell cultures, which in future could be used for identifying different lung cell types within co-cultures and studying the process of early carcinogenesis in lung cell culture.
Highlights
Raman spectroscopy is a powerful bioanalytical technique that reveals the chemical constituents of a given sample based on the inelastic scattering properties of molecular bonds
The aim of this study was to determine whether confocal Raman micro-spectroscopy would show sufficient sensitivity and specificity for identification of primary human bronchial epithelial cells (HBECs) as well as immortalized cell lines to be used for live cell biological studies in vitro
To avoid extended dwell time and allow more frequent Raman spectroscopy data acquisitions from more cells when studying primary HBECs, we examined the potential of using a line-scan rather than an area-scan data acquisition
Summary
Raman spectroscopy is a powerful bioanalytical technique that reveals the chemical constituents of a given sample based on the inelastic scattering properties of molecular bonds. The aim of this study was to determine whether confocal Raman micro-spectroscopy would show sufficient sensitivity and specificity for identification of primary human bronchial epithelial cells (HBECs) as well as immortalized cell lines to be used for live cell biological studies in vitro. To achieve our aim, we first established a detailed protocol for imaging of live human lung cells in vitro by directly comparing methods for sample preparation and data acquisition. We performed these initial optimization studies in live human lung cancer (A549) and fibroblast (MRC5) immortalized cell lines and compared the imaging results qualitatively with fluorescence imaging. May be suitable for differentiating between HBEC primary cell cultures and could in future be applied to identification of different lung cell types within co-cultures and studying the process of early lung carcinogenesis in cell culture
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
