bioRxiv ScienceSearch

Biology subjects

Samuel, A. Z.

Publications and source records attributed to Samuel, A. Z..

2 recordsLinked to original sources

Detection of Penicillin G Produced by Penicillium chrysogenum KF 425 in Vivo with Raman Microspectroscopy and Multivariate Curve Resolution-Alternating Least Squares Methods

Raman microspectroscopy is a minimally invasive technique that can identify molecular structure without labeling. In this study, we demonstrate in vivo detection of the bioactive compound penicillin G inside Penicillium chrysogenum KF425 fungus cells. Highly overlapped spectroscopic signatures acquired using Raman microspectroscopic imaging are analyzed using a multivariate curve resolution-alternating least squares (MCR-ALS) method to extract the pure spectra of individual molecular constituents. In addition to detecting multiple constituents such as proteins and lipids, we observe the subcellular localization of penicillin G like granule particle inside the fungus body. To date, there have been no reports of direct visualization of intracellular localization of penicillin G. The methodology we present in this article is expected to be applied as a screening tool for the production of bioactive compounds by microorganisms.

microbiology

Molecular profiling of lipid droplets inside HuH7 Cells with Raman micro-spectroscopy

Raman imaging has become an attractive technology in molecular biology because of its ability to detect multiple molecular components simultaneously without labeling. Two major limitations in accurately accounting for spectral features, viz. background removal and spectral unmixing, have been overcome by employing a modified and effective routine in multivariate curve resolution (MCR). With our improved strategy, we have spectrally isolated seven structurally specific biomolecules without any post-acquisition spectral treatments. Consequently, the isolated intensity profiles reflected concentrations of corresponding biomolecules with high statistical accuracy. Our study reveals the changes in the molecular composition of lipid droplets (LD) inside HuH7 cells and its relation to the physiological state of the cell. Further, we show that the accurate separation of spectral components permits analysis of in vivo structural modification of molecules after cellular uptake. A detailed discussion is presented to highlight the potential of Raman spectroscopy with MCR in semi-quantitative molecular profiling of living cells.

bioengineering