BCA deconvolutes Raman spectrum to a linear superposition of the weighted biomolecular component spectra (Proteins, DNA, RNA, Lipids, Saccharides)
Modern instrumentation for Raman microspectroscopy and current techniques in analysis of spectral data provide new opportunities to study molecular interactions and dynamics at subcellular levels in biological systems. Implementation of biomolecular component analysis (BCA) to microRaman spectrometry provides basis for the emergence of Ramanomics, a new biosensing discipline with unprecedented capabilities to measure concentrations of distinct biomolecular groups in live cells and organelles.
Raman light scattering from different types of biomolecules generates corresponding bands in the Raman spectra, which enables to resolve all the diversity of biomolecular components—nucleic acids, amino acids, proteins, and various classes of lipids and saccharides. Being applied to live cells, it provides multiple benefits for molecular analysis, including the linearity of this optical phenomenon, which allows for quantitative probing of biomolecular distribution. This makes confocal Raman spectrometry, apparently, the only in situ technique, capable of providing quantitative biochemical analysis of a single live cell.
BCA-box brings back a set of important parameters, which are usually generated as output of different omics discipline, and which are grouped here as Ramanomic parameters (concentrations of proteins, nucleic acids, saccharides, lipids and lipidome characteristics). A result of in-situ analysis by BCA of different organelles in HeLa cells – Nucleoli, Apparatus Golgi, Endoplasmic Reticulum, and Mitochondria, as an example is demonstrated. In scattered graphs each point represents single organelle Raman spectrum measurement. Automated interpretation methods organized as the BCA-box are suitable for use in a biochemistry laboratory.
HeLa cells Ramanomics
A.N.Kuzmin et al., submitted for publication
Averaged organellar concentrations of biomolecules are tabulated as a BCA analysis result. The same table can be generated either for any other cell line or for the same cell line with disease abnormality. These data can be collected in the website data base for free exchange between the scientific groups of Raman and/or biomedical community. Scattered graphs shown in previous slide can be also presented as heatmap, if necessary.
Prostate cancer diagnostics
Use BCA-box together with Confotec MR200 laser scanning Raman microscope to discover new markers of carcinogenesis. As shown in this example, Raman scattering is a promising tool for discovering new facts about cancer and processes underlying this disease.
HeLa organellar proteins analysis
For advanced users. Raman microscope Confotec MR200 combined with BCA-box provides you the ability to analyze the structure of different types of biomolecules. This slide demonstrates the results of the analysis of secondary structure motifs of proteins in different HeLa organelles.
- single organelle Raman spectrum decomposition (proteins, DNA, RNA, lipids, Glycogen concentrations);
- single Organelle lipidomics in live cell (lipid concentration, unsaturation level, trans/cis isomers ratio, level of sphingolipids andcholesterol);
- monitoring the dynamics of biological processes on a single organelle level (apoptotic cell death, RNA synthesis in nucleoli, drugintervention etc.);
- tracking the Raman and Resonance Raman probes in live cells;
- search for new markers of carcinogenesis;
- study of molecular biology of cancer;
- tracking of drug intervention in cancer cells.