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High-speed atomic force microscopy (HS-AFM) is an indispensable technique inthe biological field owing to its excellent imaging capability for thereal-time observation of biomolecules with high spatial resolution.Furthermore, recent developments have established a tip-scan stand-alone HS-AFMthat can be combined with an optical microscope, drastically improving itsversatility for studying various complex phenomena. Although HS-AFM has mainlybeen used in biology, it has considerable potential to contribute to variousresearch fields. One of the great candidates is a photoactive material, such asan azo-polymer, which plays a vital role in multiple optical applicationsbecause of its unique nanoscale motion under light irradiation. In this study,we demonstrate the in-situ real-time observation of nanoscale azo-polymermotion by combining tip-scan HS-AFM with an optical system, allowing HS-AFMobservations precisely aligned with a tightly focused laser position. Wesuccessfully observed the dynamic evolution of unique morphologies inazo-polymer films, attributed to photoinduced nano-movements. Moreover,real-time topographic line profile analyses facilitated precise andquantitative investigations of morphological changes, which provided novelinsights into the deformation mechanism. This significant demonstration wouldpave the way for the application of HS-AFM in wide research fields, frombiology to material science and physical chemistry.

In-situ real-time observation of photo-induced nanoscale azo-polymer  motions using high-speed atomic force microscopy combined with an inverted  optical microscope