Fluidic force microscopy (FluidFM), which combines atomic force microscopy (AFM) with microchanneled cantilevers connected to a pressure controller, is a technique allowing the realization of force-sensitive nanopipette under aqueous conditions. FluidFM has unique advantages in simultaneous three-dimensional manipulations and mechanical measurements of biological specimens at the micro-/nanoscale. Over the past decade, FluidFM has shown its potential in biophysical assays particularly in the investigations at single-cell level, offering novel possibilities for discovering the underlying mechanisms guiding life activities. Here, we review the utilization of FluidFM to address biomechanical and biophysical issues in the life sciences. Firstly, the fundamentals of FluidFM are represented. Subsequently, the applications of FluidFM for biophysics at single-cell level are surveyed from several facets, including single-cell manipulations, single-cell force spectroscopy, and single-cell electrophysiology. Finally, the challenges and perspectives for future progressions are provided.

Single-cell analysis has been considered as a promising way to uncover the underlying mechanisms guiding the mysteries of life activities, which considerably complements traditional ensemble assays and yields novel insights into cell biology. The advent of atomic force microscopy (AFM) provides a potent tool for investigating the structures and properties of native biological samples at the micro/nanoscale under near-physiological conditions, which promotes the studies of single-cell behaviors. In the past decades, AFM has achieved great success in single-cell observation and manipulation for biomedical applications, demonstrating the excellent capabilities of AFM in addressing biological issues at the single-cell level with unprecedented spatiotemporal resolution. In this article, we review the recent advances in single-cell analysis that has been made with the utilization of AFM, and provide perspectives for future progression.