Super-Resolution Localisation Microscopy: Resolving Fluorescence from Single Molecules in Time and Space

Dr Toby D M Bell

School of Chemistry, Monash University

3:30 pm, Friday, 16 October 2015, EN Lecture Theatre (EN Building), Hawthorn

Fluorescence is an exquisitely sensitive and extremely versatile tool for interrogating all manner of materials and is ideally suited for biological samples. In this talk the use of single molecule detection to achieve super-resolution fluorescence microscopy with spatial resolution an order of magnitude improved over the diffraction limit of light will be described including instrumentation and image rendering.[1] The importance of benchmarking and potential image artefacts and how to recognise and avoid them will also be discussed.[2] The talk will be illustrated throughout with examples of research performed in my group and with collaborators covering nanomaterials and biological systems including key cellular components such as microtubules, actin and mitochondria.[3] Recent progress in imaging microtubules in cells transfected with proteins from Rhabdovirus (the causative agent of rabies) will be presented.[4]
Single molecule super-resolution image from a COS 7 cell stained for microtubulin with Alexa647.

[1] D.R. Whelan and T.D.M. Bell, “Super-Resolution Single-Molecule Localization Microscopy: Tricks of the Trade”. J. Phys. Chem. Lett., (2015) DOI: 10.1021/jz5019702. See also: http://pubs.acs.org/page/jpclcd/bell-video.html
[2] D.R. Whelan, and T.D.M. Bell, “Correlative Synchrotron Fourier Transform Infrared Spectroscopy and Single Molecule Super Resolution Microscopy for the Detection of Composition and Ultrastructure Alterations in Single Cells”. ACS Chem. Biol., (2015), DOI: 10.1021/acschembio.5b00754
[3] D.R. Whelan, and T.D.M. Bell, “Image Artifacts in Single Molecule Localization Microscopy: Why Optimization of Sample Preparation Protocols Matters”. Sci. Rep., (2015), DOI: 10.1038/srep07924.
[4] T.D.M. Bell, D.R. Whelan, A. Brice, G.W. Moseley, “Super-Resolution Microscopy of Cells Expressing Rhabdovirus Proteins”. Biophys. J., (2014) 106, 603A.