A new approach for repeated tip-sample relocation for AFM imaging of nano and micro sized particles and cells in liquid environment.

The ability to repeatedly find exact the same nano region-of-interest (nROI) is essential for atomic force microscopy (AFM) studies of heterogeneous environmental samples. The large variety of methods makes it difficult to find the most suitable one for a specific research question. We thus conducted a literature research for nROI relocation methods and organized the found references in order to give an overview over relocation methods including the advantages, limitations and documented applications. This survey of nROI relocation methods and their key information facilitates the selection of appropriate methods with respect to a specific research question. Based on this survey, we developed a new AFM relocation approach urgently needed for the study of nano and micro sized particles and cells in air and aqueous environment. This approach uses commercially available TEM grids fully embedded in a semitransparent resin as a glue body on top of which particles and cells are fixed. Relocation of nROI within one grid is based on easily recognizable sample features in micro and nanometer scale. The stable sticking of the studied mineral particles and bacterial cells allows repeated measurements of the same nROI with differently functionalized tips in air as well as in water. Our simple, fast, and cost-effective method allows relocation with an accuracy of 10-40 nm and enables the implementation of AFM/ESEM correlative microscopy.