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.
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