New perspectives for viability studies with high-content analysis Raman spectroscopy (HCA-RS)

Abstract

Raman spectroscopy has been widely used in clinical and molecular biological studies, providing high chemical specificity without the necessity of labels and with little-to-no sample preparation. However, currently performed Raman-based studies of eukaryotic cells are still very laborious and time-consuming, resulting in a low number of sampled cells and questionable statistical validations. Furthermore, the approach requires a trained specialist to perform and analyze the experiments, rendering the method less attractive for most laboratories. In this work, we present a new high-content analysis Raman spectroscopy (HCA-RS) platform that overcomes the current challenges of conventional Raman spectroscopy implementations. HCA-RS allows sampling of a large number of cells under different physiological conditions without any user interaction. The performance of the approach is successfully demonstrated by the development of a Raman-based cell viability assay, i.e., the effect of doxorubicin concentration on monocytic THP-1 cells. A statistical model, principal component analysis combined with support vector machine (PCA-SVM), was found to successfully predict the percentage of viable cells in a mixed population and is in good agreement to results obtained by a standard cell viability assay. This study demonstrates the potential of Raman spectroscopy as a standard high-throughput tool for clinical and biological applications.

Raman Microscopy: Progress in Research on Cancer Cell Sensing

“…A similar experimental set-up was made based on a high-content analysis Raman spectroscopy (HCA-RS) platform to the development of a Raman-based cell viability assay, in terms of the effect of doxorubicin (DOX) concentration on monocytic THP-1 cells as monocytic acute myeloid leukemia cell line [ 53 ]. The drug-induced changes in cells were then investigated by HCA-RS, with, in total, 25,031 cells sampled to differentiate between viable and non-viable cells by PCA combined with support vector machine (PCA-SVM).…”

Probing metabolic alterations in breast cancer in response to molecular inhibitors with Raman spectroscopy and validated with mass spectrometry

“…[23][24][25] RS is also nondestructive allowing sample archival and retesting for accurate measure of therapeutic response. [26][27][28][29] Here, we leveraged the advantages of Raman spectral mapping in its ability to rst measure dynamic changes at the single-cell level with high sensitivity; second spatiotemporally resolve multiplexed metabolic changes; and third enable quantitative analysis. We treated triple-negative BC cell line MDA-MB-231 with Trametinib, a potent and specic MEK1/2 allosteric inhibitor, 30 that downregulates MEK signaling in the ERK pathway.…”

Leukocyte Activation Profile Assessed by Raman Spectroscopy Helps Diagnosing Infection and Sepsis

“…Furthermore, as Raman spectroscopy is based on fast (instantaneous) light-matter interactions, the method has the potential to provide the result within the recommended “Golden Hour” for the diagnosis of sepsis. Recently, Raman technology has advanced to high-throughput devices which enable the analysis of greater than 1,000 cells within only 1 hour ( 19 , 43 ) paving the way for larger, multicenter clinical trials to validate spectroscopic immunophenotyping. This will enable to overcome the limitation of the current study and to include more patients, so that the impact of timing of sampling, type of pathogen as well as age, sex, ethnicity, or race of the patient can be fully assessed.…”