by André Körnig, JPK BioAFM, Bruker Nano GmbH
NanoWizard® V BioScience – The latest Generation AFM for Automated Structural and Mechanical Analysis & NanoRacer® – High-Speed AFM for dynamical processes
The ability of atomic force microscopy (AFM) to obtain three-dimensional topography images of biological molecules and complexes with nanometer resolution and under near-physiological conditions remains unmatched by other imaging techniques. Bruker BioAFM has recently launched the NanoWizard® V BioScience AFM that combines high spatio-temporal resolution with a large scan area, flexible experiment design, and outstanding integration with advanced optical microscope systems.
This talk will focus on how the advances in Bruker’s latest BioAFM can be applied to study a wide-range of biological samples: from individual biomolecules to mammalian cells and tissues in-situ. It will be presented how we are able to resolve the nanoscale structure of individual biomolecules, at high-speed scan rates (400 lines/sec), follow the dynamic reorganization of the membrane-associated cytoskeleton of living cells at high temporal and spatial resolution. It will be highlighted, how the topography of cells across the entire area of the microscope stage can be automatically mapped. Special part will be dedicated to the suite of BioAFM modes, probes and accessories for studying nanomechanical properties of cells and tissues, including direct correlation with super-resolution microscopy techniques (STED).
In the past, investigating large and rough samples such as tissues and hydrogels using AFM was challenging due to the limited z-axis of the AFM. Using osteoarthritic cartilage as an example, we will demonstrate how a newly developed hybrid of a motorized and piezo stage enables multi-region AFM probing over a large, rough sample area while providing additional correlative optical data sets.
The newly developed NanoRacer® High-Speed AFM enables scanning speeds of up to 50 frames per second. In this way, the high-speed study of the time-resolved dynamics associated with cellular processes and the binding mechanisms of individual biomolecules is possible, e.g. the dynamics of single molecule binding behavior, two-dimensional protein assemblies, motor proteins and membrane trafficking.
We will present data on DNA origami nanostructures containing biotin binding sites, imaged in fluid in the presence of streptavidin, as well as data on DNA metastable bubble formation and closure (~30 nm in length), imaged in fluid in closed-loop at 2000 lines per second.