A software tool for STED-AFM correlative super-resolution microscopy

Sami Koho; Takahiro Deguchi; Madis Löhmus; Tuomas Näreoja; Pekka E. Hänninen

doi:10.1117/12.2078624

9 March 2015 A software tool for STED-AFM correlative super-resolution microscopy

Sami Koho, Takahiro Deguchi, Madis Löhmus, Tuomas Näreoja, Pekka E. Hänninen

Proceedings Volume 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII; 93301K (2015) https://doi.org/10.1117/12.2078624
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

Multi-modal correlative microscopy allows combining the strengths of several imaging techniques to provide unique contrast. However it is not always straightforward to setup instruments for such customized experiments, as most microscope manufacturers use their own proprietary software, with limited or no capability to interface with other instruments - this makes correlation of the multi-modal data extremely challenging. We introduce a new software tool for simultaneous use of a STimulated Emission Depletion (STED) microscope with an Atomic Force Microscope (AFM). In our experiments, a Leica TCS STED commercial super-resolution microscope, together with an Agilent 5500ilm AFM microscope was used. With our software, it is possible to synchronize the data acquisition between the STED and AFM instruments, as well as to perform automatic registration of the AFM images with the super-resolution STED images. The software was realized in LabVIEW; the registration part was also implemented as an ImageJ script. The synchronization was realized by controlling simple trigger signals, also available in the commercial STED microscope, with a low-cost National Instruments USB-6501 digital I/O card. The registration was based on detecting the positions of the AFM tip inside the STED fieldof-view, which were then used as registration landmarks. The registration should work on any STED and tip-scanning AFM microscope combination, at nanometer-scale precision. Our STED-AFM correlation method has been tested with a variety of nanoparticle and fixed cell samples. The software will be released under BSD open-source license.

Citation Download Citation

Sami Koho, Takahiro Deguchi, Madis Löhmus, Tuomas Näreoja, and Pekka E. Hänninen "A software tool for STED-AFM correlative super-resolution microscopy", Proc. SPIE 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII, 93301K (9 March 2015); https://doi.org/10.1117/12.2078624

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