Combining AFM with Super-Resolution STED (Simulated Emission Depletion) Microscopy

_{KEYWORDS: Atomic Force Microscopy; AFM; NanoWizard; Super-resolution; STED; Correlative Microscopy; Cell Biology; Life Science}

Research in life sciences demands more and more from the analysis capabilities of biological samples with the highest resolution. This requires the development of new imaging techniques, both optical and non-optical. Non-invasive instrumentation such as atomic force microscopy AFM) provides high resolution topography without requiring labeling of the sample. The label-free topographical, electrical, magnetic, and mechanical information gained from the AFM correlated with the color-encoded molecular fluorescence map provides an important tool for deeper investigation of sample properties. When AFM is combined with a super-resolution microscopy system, such as stimulated emission depletion (STED), it generates a nanoscopic tool with high versatility to answer sub-diffraction level morphology questions.

This application note exemplifies how a system integration of AFM and STED works. The data included in this app note have been reported in a paper published by the group of Prof. Alberto Diaspro from IIT in Genova, Italy in the journal, Optical Nanoscopy (https://doi.org/10.1016/j.bbamem.2011.07.024).

Readers can expect to learn about:

• Basic principle of an AFM and STED based system;
• Prominent application data showcasing the versatility and the seamless integration of Bruker's NanoWizard AFM into advanced optical microscopes; and
• The potential of a combined AFM and STED system for studying biological processes such as vesicle movement, dendritic spine activity or actin dynamics within living cells.