A Correlative SICM‐OPM Platform for Surface and Volumetric Imaging in Live Cells

ABSTRACT Live‐cell imaging of cell surface topography and intracellular architecture is essential for understanding cellular function. However, conventional approaches often involve trade‐offs between resolution, invasiveness, and volumetric coverage. Here, we present an integrated Scanning Ion Conductance Microscope and single‐objective Oblique Plane Microscope (SICM‐OPM) system that enables simultaneous non‐contact topographical imaging and volumetric fluorescence imaging within the same live cell without sample translation. Beyond correlative live imaging, the platform supports nanomechanical mapping with tens‐of‐nanometers resolution, fluorescence‐guided localized molecular delivery via the SICM, and benefits from reduced photobleaching due to light‐sheet excitation. We demonstrate this platform's capabilities by visualizing imipramine‐induced transverse‐tubule (TT) remodeling in live cardiomyocytes, revealing pronounced detubulation of internal TT invaginations while surface TT opening characteristics remain largely preserved, and capturing high‐speed correlative volumetric images of clathrin‐mediated endocytosis in Cos‐7 cells. Additionally, we show precision delivery of fluorescent cargos, including dextrans and α‐synuclein, into mammalian cells and diatoms, alongside localized stiffness mapping to evaluate mechanical responses of mammalian cells. We believe this technique opens new avenues for correlative structural, functional, and biophysical studies in live cells, with broad relevance to cell biology, neurodegeneration, and mechanobiology.