In the following study, we demonstrate the power of combining atomic force microscopy AFM and fluorescence microscopy techniques to probe real-time, in-situ effects of two highly specific drugs that are able to disrupt different cytoskeleton networks inside living cells. Using Veeco Multiple Image Registration Overlay (MIRO) software and the new Bioscope Catalyst Atomic Force Microscope (Veeco Instruments, Santa Barbara, CA), we realized unprecedented accuracy in terms of navigation and force curves quality, establishing this important new imaging technique as a foundation for many exciting applications in cancer research.