Publications by Seamus John Holden

Super-resolution fight club: assessment of 2D and 3D single-molecule localization microscopy software

Michaël Unser, Daniel Sage, Ricardo Henriques, Thanh-An Michel Pham, Thomas Pengo, Seamus John Holden, Tomas Lukes, Anna Archetti, Silvia Colabrese, Anurag Agrawal

With the widespread uptake of two-dimensional (2D) and three-dimensional (3D) single-molecule localization microscopy (SMLM), a large set of different data analysis packages have been developed to generate super-resolution images. In a large community effo ...

2019

Constriction Rate Modulation Can Drive Cell Size Control and Homeostasis in C. crescentus

Suliana Manley, Seamus John Holden, Aster Louis K Vanhecke, Anna Archetti, Ambroise Justin Lambert, Felix Max Schaber

Rod-shaped bacteria typically grow first via sporadic and dispersed elongation along their lateral walls and then via a combination of zonal elongation and constriction at the division site to form the poles of daughter cells. Although constriction compris ...

2018

Functional dichotomy and distinct nanoscale assemblies of acell cycle-c ontrolled bipolar zinc-finger regulator

Suliana Manley, Seamus John Holden

Protein polarization underlies differentiation in metazoans and in bacteria. How symmetric polarization can instate functional asymmetry remains elusive. Here, we show by super-resolution photo-activated localization microscopy and edgetic mutations that t ...

Elife Sciences Publications Ltd2016

Modularity and determinants of a (bi-)polarization control system from free-living and obligate intracellular bacteria

Suliana Manley, Seamus John Holden

Although free-living and obligate intracellular bacteria are both polarized it is unclear whether the underlying polarization mechanisms and effector proteins are conserved. Here we dissect at the cytological, functional and structural level a conserved po ...

Elife Sciences Publications Ltd2016

PALMsiever: a tool to turn raw data into results for single-molecule localization microscopy

Suliana Manley, Thomas Pengo, Seamus John Holden

During the past decade, localization microscopy (LM) has transformed into an accessible, commercially available technique for life sciences. However, data processing can be challenging to the non-specialist and care is still needed to produce meaningful re ...

Oxford University Press2015

Correction of a Depth-Dependent Lateral Distortion in 3D Super-Resolution Imaging

Suliana Manley, Seamus John Holden, Lina Carlini, Kyle Michael Douglass

Three-dimensional (3D) localization-based super-resolution microscopy (SR) requires correction of aberrations to accurately represent 3D structure. Here we show how a depth-dependent lateral shift in the apparent position of a fluorescent point source, whi ...

Public Library of Science2015

3D high-density localization microscopy using hybrid astigmatic/biplane imaging and sparse image reconstruction

Michaël Unser, Suliana Manley, Seamus John Holden, Lina Carlini

Localization microscopy achieves nanoscale spatial resolution by iterative localization of sparsely activated molecules, which generally leads to a long acquisition time. By implementing advanced algorithms to treat overlapping point spread functions (PSFs ...

Optical Soc Amer2014

High throughput 3D Palm Imaging Elucidates Mechanisms of Bacterial Cell Division

Suliana Manley, Thomas Pengo, Seamus John Holden

Cell Press2014

FALCON: fast and unbiased reconstruction of high-density super-resolution microscopy data

Michaël Unser, Cédric René Jean Vonesch, Suliana Manley, Hagay Kirshner, Seamus John Holden, Lina Carlini

Super resolution microscopy such as STORM and (F) PALM is now a well known method for biological studies at the nanometer scale. However, conventional imaging schemes based on sparse activation of photo-switchable fluorescent probes have inherently slow te ...

Nature Publishing Group2014

Super-Resolution Fluorescence Microscopy of Transcription Sites in E. coli

Seamus John Holden

Cell Press2014

High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization

Suliana Manley, Thomas Pengo, Seamus John Holden, Karin Lederballe Meibom

We created a high-throughput modality of photoactivated localization microscopy (PALM) that enables automated 3D PALM imaging of hundreds of synchronized bacteria during all stages of the cell cycle. We used high-throughput PALM to investigate the nanoscal ...

National Academy of Sciences2014

Multiscale Spatial Organization of RNA Polymerase in Escherichia coli

Seamus John Holden

Nucleic acid synthesis is spatially organized in many organisms. In bacteria, however, the spatial distribution of transcription remains obscure, owing largely to the diffraction limit of conventional light microscopy (200-300 nm). Here, we use photoactiva ...

Cell Press2013