Abstract
Raman microspectroscopy is a rapidly developing technique, which has an unparalleled potential for in situ proteomics, lipidomics, and metabolomics, due to its remarkable capability to analyze the molecular composition of live cells and single cellular organelles. However, the scope of Raman spectroscopy for bio-applications is limited by a lack of software tools for express-analysis of biomolecular composition based on Raman spectra. In this study, we have developed the first software toolbox for immediate analysis of intracellular Raman spectra using a powerful biomolecular component analysis (BCA) algorithm. Our software could be easily integrated with commercial Raman spectroscopy instrumentation, and serve for precise analysis of molecular content in major cellular organelles, including nucleoli, endoplasmic reticulum, Golgi apparatus, and mitochondria of either live or fixed cells. The proposed software may be applied in broad directions of cell science, and serve for further advancement and standardization of Raman spectroscopy.
Highlights
Raman spectroscopy and microscopy, in particular, holds its unique niche in the ever-expanding family of biosensing tools, bridging conventional microscopic imaging together with a molecular analysis of the samples
2018, 8,range, 4 of 11of wavelength with standard error produced by the Raman microscope during measurement egg white protein mixture (±3 counts per second, cps)
Results and Discussion wavelength range, with standard error produced by the Raman microscope during measurement of egg protein mixture
Summary
In particular, holds its unique niche in the ever-expanding family of biosensing tools, bridging conventional microscopic imaging together with a molecular analysis of the samples. This optical technique utilizes inelastic light scattering from the molecular bond vibrations, which produce their characteristic Raman spectral bands, enabling selective detection of diverse biomolecules in a sample that includes certain amino acids, DNA, and RNA, as well as various classes of lipids, proteins, and saccharides. Maturation of the Raman spectroscopic technology leads to an emergence of Ramanomics, an independent “Omics” discipline, aimed at quantitative characterization and monitoring of molecular content in biological samples [8]
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