Sunday , February 21 , 2010 179 a

925-Pos Novel Visualisation Techniques for Localisation Microscopy David Baddeley, Isuru D. Jayasinghe, Sabrina Roßberger, Mark B. Cannell, Christian Soeller. University of Auckland, Auckland, New Zealand. Localisation microscopy techniques based on localising single fluorophore molecules now routinely achieve accuracies better than 30 nm. Unlike conventional optical microscopy approaches, localisation microscopy experiments do not generate an image but a list of discrete coordinates of estimated fluorophore positions, typically involving 10,000 to 100,000 molecule coordinates. Data display and analysis therefore generally requires visualisation methods that translate the position data into conventional images. Here we investigate the properties of several widely used visualisation techniques and show that a commonly used algorithm based on rendering Gaussians may lead to a 1.44-fold loss of resolution. Additionally, existing methods typically do not explicitly take sampling considerations into account and thus may produce spurious structures. To overcome some of these issues we present two additional visualisation algorithms, an adaptive histogram method based on quad trees and a Delaunay triangulation based visualisation of point data. The new visualisation methods are designed to supress erroneous detail in poorly sampled image areas but avoid loss of resolution in well sampled regions. A number of criteria for scoring visualisation methods are developed as a guide for choosing among visualisation methods and are used to qualitatively compare various algorithms. We show how these algorithms can be extended to visualise 3D localisation data and demonstrate in practical cell labelling experiments that the effective resolution is typically sampling-limited. The visualisation techniques are illustrated with 2D and 3D localisation data obtained in cardiac ventricular myocytes stained for caveolin-3, ryanodine receptors and b-tubulin.