title: Reduction of coherent artefacts in super-resolution fluorescence localisation microscopy.
abstract: Super-resolution localisation microscopy techniques depend on uniform illumination across the field of view, otherwise the resolution is degraded, resulting in imaging artefacts such as fringes. Lasers are currently the light source of choice for switching fluorophores in PALM/STORM methods due to their high power and narrow bandwidth. However, the high coherence of these sources often creates interference phenomena in the microscopes, with associated fringes/speckle artefacts in the images. We quantitatively demonstrate the use of a polymer membrane speckle scrambler to reduce the effect of the coherence phenomena. The effects of speckle in the illumination plane, at the camera and after software localisation of the fluorophores, were characterised. Speckle phenomena degrade the resolution of the microscope at large length scales in reconstructed images, effects that were suppressed by the speckle scrambler, but the small length scale resolution is unchanged at ∼30 nm.
authors.affiliation: Biological Physics, School of Physics and Astronomy, The University of Manchester, Manchester, U.K.|Faculty of Life Sciences, Michael Smith Building, The University of Manchester, Oxford Road, Manchester, M13 9PT, U.K.
authors.forename: Pantelis
authors.initials: P
authors.lastname: Georgiades
authors.affiliation: Faculty of Life Sciences, Michael Smith Building, The University of Manchester, Oxford Road, Manchester, M13 9PT, U.K.|Photon Science Institute, The University of Manchester, Oxford Road, Manchester, M13 9PL, U.K.
authors.forename: Viki J
authors.initials: VJ
authors.lastname: Allan
authors.affiliation: Photon Science Institute, The University of Manchester, Oxford Road, Manchester, M13 9PL, U.K.
authors.forename: Mark
authors.initials: M
authors.lastname: Dickinson
authors.affiliation: Biological Physics, School of Physics and Astronomy, The University of Manchester, Manchester, U.K.|Photon Science Institute, The University of Manchester, Oxford Road, Manchester, M13 9PL, U.K.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Waigh
keywords: Fluorescence; PALM; STORM; TIRF; speckle; super-resolution