title: PALM and STORM: Into large fields and high-throughput microscopy with sCMOS detectors.
abstract: Single Molecule Localization Microscopy (SMLM) techniques such as Photo-Activation Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) enable fluorescence microscopy super-resolution: the overcoming of the resolution barrier imposed by the diffraction of light. These techniques are based on acquiring hundreds or thousands of images of single molecules, locating them and reconstructing a higher-resolution image from the high-precision localizations. These methods generally imply a considerable trade-off between imaging speed and resolution, limiting their applicability to high-throughput workflows. Recent advancements in scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera sensors and localization algorithms reduce the temporal requirements for SMLM, pushing it toward high-throughput microscopy. Here we outline the decisions researchers face when considering how to adapt hardware on a new system for sCMOS sensors with high-throughput in mind.
authors.affiliation: Quantitative Imaging and NanoBiophysics Group, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, WC1E 6BT London, United Kingdom.
authors.forename: Pedro
authors.initials: P
authors.lastname: Almada
authors.affiliation: Quantitative Imaging and NanoBiophysics Group, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, WC1E 6BT London, United Kingdom.
authors.forename: Siân
authors.initials: S
authors.lastname: Culley
authors.affiliation: Quantitative Imaging and NanoBiophysics Group, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, WC1E 6BT London, United Kingdom. Electronic address: r.henriques@ucl.ac.uk.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Henriques
mesh_terms: D000465:Algorithms; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D010784:Photomicrography / Q000295:instrumentation / Q000379:methods*
keywords: Hardware; Homogenization; Single-molecule localization; sCMOS