Sponsored Article: Correlative Nanoscopy - AFM-STED Combination

Quantitative and fast Atomic force microscopy (AFM) co-localized with STED microscopy for correlative experiments on living cells

Atomic force microscopy (AFM) has emerged as a powerful tool for life science research. It can be used to investigate a huge variety of samples with nanometer resolution under physiological conditions. In addition to delivering topographic measurements, it enables the determination of elasticity, adhesion and other nanomechanical characteristics and the high-speed study of the time-resolved dynamics associated with cellular processes. Modes like PeakForce Tapping™ and QI™ mode combined with easy-to-use features allow even beginners to create high quality data of topography, adhesion and mechanical information.

Thanks to its tip-scanning design, Bruker BioAFM’s are perfectly designed to combine with optical microscopes. Fast AFM imaging of several frames per second can be seamlessly combined with methods such as epifluorescence, confocal, TIRF, STED microscopy. The combination of AFM and optical/fluorescence microscopy offers the ideal way to gain access to chemical recognition and to extend the capabilities of AFM.

In the past decade, super-resolution light microscopy has attracted significant attention in the scientific community. Stimulated Emission Depletion (STED) microscopy enables sub-diffraction limit resolution, and new lasers and concepts like easySTED have allowed the development of robust super-resolution microscopes. Atomic force microscopy and STED microscopy address a similar nanometer resolution in biological samples. STED microscopy provides molecular specificity and intracellular information while AFM delivers true surface visualization in 3D and nanomechanical information.

The combination and overlay of AFM and STED images makes it possible to create a more complete set of data, correlate different information and gain a more complex understanding of the underlying features and characteristics. The combined approach of AFM manipulation and STED microscopy enables the correlation of information on chemical recognition, intracellular features and topography/nanomechanical information with nanometer resolution, and is a very powerful tool for investigating cells and biological specimen.

Bruker’s patented DirectOverlay™ feature enables perfect optical integration. It allows the precise correlation of AFM and optical data by performing a perfect overlay of the data sets and overcomes any mismatch caused by errors in the optical lens systems.

The performance, reliability and benefits of a combined AFM and STED show that correlative nanoscopy is a powerful tool for biological applications.

For more information, please visit www.bruker.com/bioafm or contact us at [email protected]

Combined AFM-STED on living human skin fibroblast:
A) Microtubules labelled with SiR Tubulin and imaged with Abberior Instruments Expert Line STED
B) 3D topography AFM image, 12.5μm x 9μm; range: 1μm
C) 3D topography image overlaid with Young’s modulus range: 60kPa

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