Simultaneous observation of cytotoxicity and drug distribution at the effector site will bring new insights into understanding mechanism of action of drug. Here, we demonstrated the application of Raman microscopy to visualize drug-induced cytotoxicity and drug distribution as well as its mechanism of action at a real-time and single-cell level. The drug concentration and treatment-dependent decrease of reduced cyt c was found as an early marker of cell apoptosis which is more sensitive than the conventional cell viability assay. This method allows for us to screen the drug cytotoxicity and approach the mechanism at the same time, thereby reducing the cost and time of drug screening.
Amino acids and peptides are basic components of proteins and have vital importance in various biological functions and diseases. In this research, we have attempted to detect and distinguish 20 kinds of amino acids and 39 kinds of peptides without any labeling. By using Raman microscopy, more than two thousand Raman spectra were obtained within five minutes from each analyte, at femtomolecular levels. Furthermore, deep learning analyses of the spectra yielded accuracies greater than 96 percent in discriminating between the amino acids and the peptides.
Raman microscopy is a powerful tool to observe molecular distribution in live cells. Here, we propose a technique to detect small molecule drugs using alkyne-tag surface-enhanced Raman scattering (SERS) imaging. To obtain SERS effect, we use gold nanoparticles as SERS probes so that we can enhance Raman scattering of small molecule drugs at low concentration with several orders of magnitude. We use alkyne tag to selectively detect the drug molecules by using the Raman peak of alkyne in the spectral silent region. Home-built slit-scanning Raman microscopy enables us to perform rapid SERS imaging. We successfully detect SERS signal from an alkyne-tagged inhibitor of a lysosomal enzyme with gold nanoparticle modified glass substrate.
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