Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data.

J Microsc

Department of Mechanical Engineering, University of California, Santa Barbara, California, U.S.A.

Published: March 2018

Modern fluorescence microscopy enables fast 3D imaging of biological and inert systems alike. In many studies, it is important to detect the surface of objects and quantitatively characterize its local geometry, including its mean curvature. We present a fully automated algorithm to determine the location and curvatures of an object from 3D fluorescence images, such as those obtained using confocal or light-sheet microscopy. The algorithm aims at reconstructing surface labelled objects with spherical topology and mild deformations from the spherical geometry with high accuracy, rather than reconstructing arbitrarily deformed objects with lower fidelity. Using both synthetic data with known geometrical characteristics and experimental data of spherical objects, we characterize the algorithm's accuracy over the range of conditions and parameters typically encountered in 3D fluorescence imaging. We show that the algorithm can detect the location of the surface and obtain a map of local mean curvatures with relative errors typically below 2% and 20%, respectively, even in the presence of substantial levels of noise. Finally, we apply this algorithm to analyse the shape and curvature map of fluorescently labelled oil droplets embedded within multicellular aggregates and deformed by cellular forces.

Download full-text PDF

Source
http://dx.doi.org/10.1111/jmi.12624DOI Listing

Publication Analysis

Top Keywords

fluorescently labelled
8
geometrical characterization
4
characterization fluorescently
4
labelled surfaces
4
surfaces noisy
4
noisy microscopy
4
microscopy data
4
data modern
4
modern fluorescence
4
fluorescence microscopy
4

Similar Publications

Aptamer-based fluorescence biosensor for rapid detection of chloramphenicol based on pyrene excimer switch.

Anal Bioanal Chem

January 2025

State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China.

Chloramphenicol (CAP) is widely used in treating bacteria infection in animals and humans. However, the accumulation of CAP in food and environment caused serious health risk to human. Consequently, sensitive and selective detection of CAP is of great importance in environmental monitoring and food safety.

View Article and Find Full Text PDF

Green fluorescent protein (GFP) chromophores are widely studied as fluorescent moieties for sensing and imaging applications. Herein, we present a straightforward synthetic strategy that involves the reaction of glycine amides with 1,3-diketones to form imidazolones through an unusual molecular fragmentation and recombination pathway. Mechanistic investigations, including crossover experiments, inspired a competing strategy that incorporates exogenous ketones into the products, yielding fluorescent GFP chromophore analogues.

View Article and Find Full Text PDF

The reactivation of heterotrimeric protein phosphatase 2A (PP2A) through small molecule activators is of interest to therapeutic intervention due to its dysregulation, which is linked to chronic conditions. This study focuses on the PP2A scaffold subunit PR65 and a small molecule activator, ATUX-8385, designed to bind directly to this subunit. Using a label-free single-molecule approach with nanoaperture optical tweezers (NOT), we quantify its binding, obtaining a dissociation constant of 13.

View Article and Find Full Text PDF

Alzheimer's disease (AD) is an age-related neurodegenerative pathology. Brain-derived extracellular vesicles (EVs) have been demonstrated to be implicated in AD pathogenesis by facilitating the propagation of Tau, amyloid-β and inflammatory cytokines. However, the impact of peripheral EVs (pEVs) in AD pathogenesis remains poorly investigated.

View Article and Find Full Text PDF

Retroviruses are responsible for significant pathology in humans and animals, including the acquired immunodeficiency syndrome and a wide range of malignancies. A crucial yet poorly understood step in the replication cycle is the recognition and selection of unspliced viral RNA (USvRNA) by the retroviral Gag protein, which binds to the psi (Ψ) packaging sequence in the 5' leader, to package it as genomic RNA (gRNA) into nascent virions. It was previously thought that Gag initially bound gRNA in the cytoplasm.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!