What you see here is a super resolution image of cos7 cells with alpha tubulin labeled with immunostaining. The second image is a zoomed area (white frame) from the first one. The raw data has been acquired at the Max Planck Institute of Biochemistry (Jungmann Group) with a pco.panda 4.2 camera.
Courtesy of Max Planck Institute of Biochemistry, Prof. Dr. R. Jungmann, A. Auer et al (manuscript in submission)
Single-Molecule Kinetics and Super-Resolution Microscopy by Fluorescence Imaging of Transient Binding on DNA Origami
DNA origami is a powerful method for the programmable assembly of nanoscale molecular structures. For applications of these structures as functional biomaterials, the study of reaction kinetics and dynamic processes in real time and with high spatial resolution becomes increasingly important. We present a single-molecule assay for the study of binding and unbinding kinetics on DNA origami. We find that the kinetics of hybridization to single-stranded extensions on DNA origami is similar to isolated substrateimmobilized DNA with a slight position dependence on the origami. On the basis of the knowledge of the kinetics, we exploit reversible specific binding of labeled oligonucleotides to DNA nanostructures for PAINT (points accumulation for imaging in nanoscale topography) imaging with <30 nm resolution. The method is demonstrated for flat monomeric DNA structures as well as multimeric, ribbon-like DNA structures.
Ralf Jungmann, Christian Steinhauer, Max Scheible, Anton Kuzyk, Philip Tinnefeld and Friedrich C. Simmel, 2010.
Lehrstuhl für Bioelektronik, Physik-Department, Technische Universität München; Angewandte Physik-Biophysik, Ludwig-Maximilians-Universität; Center for NanoScience, Ludwig-Maximilians-Universität; Physikalische und Theoretische Chemie - NanoBioScience, Technische Universität Braunschweig, Germany