Thomas Pengo

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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 nanoscale organization of the bacterial cell division protein FtsZ in live Caulobacter crescentus. We observed(More)
The HIV structural protein Gag assembles to form spherical particles of radius ∼70 nm. During the assembly process, the number of Gag proteins increases over several orders of magnitude from a few at nucleation to thousands at completion. The challenge in studying protein assembly lies in the fact that current methods such as standard fluorescence or(More)
The quality of super-resolution images obtained by single-molecule localization microscopy (SMLM) depends largely on the software used to detect and accurately localize point sources. In this work, we focus on the computational aspects of super-resolution microscopy and present a comprehensive evaluation of localization software packages. Our philosophy is(More)
The ample variety of labeling dyes and staining methods available in fluorescence microscopy has enabled biologists to advance in the understanding of living organisms at cellular and molecular level. When two or more fluorescent dyes are used in the same preparation, or one dye is used in the presence of autofluorescence, the separation of the fluorescent(More)
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 results. PALMsiever has been developed to provide a user-friendly means of visualizing, filtering and(More)
Super-resolution localization microscopy methods such as PALM and STORM have been shown to provide imaging with resolutions up to a few tens of nanometers while using relatively simple setups. Biplane PALM has extended the PALM technique to three-dimensions, by simultaneously using two imaging planes, with different focal depths. A key aspect in achieving(More)
We present and validate a quantitative, multidimensional image analysis protocol to assist in the early detection of lung cancer in minimal samples of bronchoalveolar lavage (BAL). To that end we stained BAL samples using Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms (FICTION). Our method allows(More)
Lung cancer is the deadliest form of cancer mainly because of the absence of reliable early diagnostic protocols. Therefore, there is increasing interest in the development of novel diagnostic noninvasive technologies that may improve the early detection of the disease. Bronchoscope-guided bronchoalveolar lavage (BAL) is a minimally invasive diagnostic(More)
In the localisation stage, the two stacks are processed separately. Local maxima are assumed to originate from fluorophore beads, and a threshold value is then used to keep the prominent ones only [2]. Each one of these local maxima is then fitted with a PSF model, yielding the lateral and the axial coordinates of each fluorophore bead. The fitting error is(More)
We present the hardware and software specification of a quantitative, multidimensional and multispectral microscopy system designed for the detection of lung cancer using minimal samples of bronchoalveolar lavage (BAL). BAL samples were stained using FICTION: Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of(More)
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