Advances in scanning probe microscopy for biological systems
image: Schematic diagram of the working principles of STM and AFM. A Schematic diagram of the STM system (Yang and Freund 2024). B,C STM constant current mode imaging (B) and constant height mode imaging (C) (Mironov 2014). D Schematic diagram of AFM (Voigtländer 2015). E−G Contact mode of AFM (E), tapping mode of AFM (F), and non-contact mode of AFM (G) (McKay 2021). H Force spectroscopy mode of AFM (Liu and Wang 2010)
Credit: HIGHER EDUCATION PRESS
Scanning Probe Microscopy (SPM), as a nanoscale characterization technique, employs a sharp probe to detect local tip-sample interactions through near-field physical phenomena. This approach achieves atomic-resolution surface imaging while enabling concurrent characterization of multi-parametric properties—electrical, magnetic, and chemical signals. This review offers a cross-disciplinary perspective on the advances in SPM for biological systems, which serves as a practical guide for life scientists to select from the expanding array of SPM techniques. We outline the fundamental principles of Scanning Tunneling Microscopy (STM) and Atomic Force Microscopy (AFM), before discussing a series of advanced SPM techniques: force spectroscopy for nanomechanical characterization, Kelvin Probe Force Microscopy (KPFM) for surface potential imaging, Scanning Near-field Optical Microscopy (SNOM) for super-resolution optics, Tip-Enhanced Raman Spectroscopy (TERS) for nanoscale chemical identification, and Scanning Electrochemical Microscopy (SECM) for localized electrochemical activity detection. A systematic comparison of these technologies provides researchers with clear criteria to select the optimal methodology for diverse demands, either characterizing nucleic acids and proteins or analyzing single-cell ultrastructure and biomechanics. In addition, this review explores the transformative integration of SPM and Artificial Intelligence (AI). This integration is expected to automate SPM workflows. It will also increase the stability of SPM systems and enhance the reproducibility of experimental results. Furthermore, by addressing current challenges and future perspectives of in vivo imaging, this review aims not merely to review the progress but to empower biologists to harness these intelligent multi-modal SPM systems for groundbreaking discoveries.
The work entitled “Advances in scanning probe mcroscopy for biological systems” was published in Biophysics Reports (ahead of print in Aug. 2026).