PSF identification applied to 3D fluorescence microscopy quantification

2001 ◽  
Author(s):  
Alain Dieterlen ◽  
Marie-Pierre Gramain ◽  
Chengqi Xu ◽  
Francois H. Guillemin ◽  
Serge Jacquey
Keyword(s):  
Fluorescence Microscopy ◽  
3D Fluorescence Microscopy ◽  
Psf Identification

scholarly journals Fast Segmentation From Blurred Data in 3D Fluorescence Microscopy

2017 ◽  
Vol 26 (10) ◽  
pp. 4856-4870 ◽  
Author(s):  
Martin Storath ◽  
Dennis Rickert ◽  
Michael Unser ◽  
Andreas Weinmann
Keyword(s):  
Fluorescence Microscopy ◽  
3D Fluorescence Microscopy

scholarly journals A dynamic, ring-forming MucB / RseB-like protein influences spore shape in Bacillus subtilis

PLoS Genetics ◽  
2020 ◽  
Vol 16 (12) ◽  
pp. e1009246
Author(s):  
Johana Luhur ◽  
Helena Chan ◽  
Benson Kachappilly ◽  
Ahmed Mohamed ◽  
Cécile Morlot ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Fluorescence Microscopy ◽  
Mother Cell ◽  
Morphological Changes ◽  
Cell Envelope ◽  
Genetic Relationships ◽  
Protein Domain ◽  
Phenotypic Characterization ◽  
Spore Coat ◽  
3D Fluorescence Microscopy

How organisms develop into specific shapes is a central question in biology. The maintenance of bacterial shape is connected to the assembly and remodelling of the cell envelope. In endospore-forming bacteria, the pre-spore compartment (the forespore) undergoes morphological changes that result in a spore of defined shape, with a complex, multi-layered cell envelope. However, the mechanisms that govern spore shape remain poorly understood. Here, using a combination of fluorescence microscopy, quantitative image analysis, molecular genetics and transmission electron microscopy, we show that SsdC (formerly YdcC), a poorly-characterized new member of the MucB / RseB family of proteins that bind lipopolysaccharide in diderm bacteria, influences spore shape in the monoderm Bacillus subtilis. Sporulating cells lacking SsdC fail to adopt the typical oblong shape of wild-type forespores and are instead rounder. 2D and 3D-fluorescence microscopy suggest that SsdC forms a discontinuous, dynamic ring-like structure in the peripheral membrane of the mother cell, near the mother cell proximal pole of the forespore. A synthetic sporulation screen identified genetic relationships between ssdC and genes involved in the assembly of the spore coat. Phenotypic characterization of these mutants revealed that spore shape, and SsdC localization, depend on the coat basement layer proteins SpoVM and SpoIVA, the encasement protein SpoVID and the inner coat protein SafA. Importantly, we found that the ΔssdC mutant produces spores with an abnormal-looking cortex, and abolishing cortex synthesis in the mutant largely suppresses its shape defects. Thus, SsdC appears to play a role in the proper assembly of the spore cortex, through connections to the spore coat. Collectively, our data suggest functional diversification of the MucB / RseB protein domain between diderm and monoderm bacteria and identify SsdC as an important factor in spore shape development.

scholarly journals Precise quantification of silica and ceria nanoparticle uptake revealed by 3D fluorescence microscopy

2014 ◽  
Vol 5 ◽  
pp. 1616-1624 ◽  
Author(s):  
Adriano A Torrano ◽  
Christoph Bräuchle
Keyword(s):  
Endothelial Cells ◽  
Fluorescence Microscopy ◽  
Silica Nanoparticles ◽  
Vascular Endothelial Cells ◽  
Cancer Cell Line ◽  
Absolute Number ◽  
Cervix Carcinoma ◽  
Precise Quantification ◽  
3D Fluorescence Microscopy ◽  
The Impact

Particle_in_Cell-3D is a powerful method to quantify the cellular uptake of nanoparticles. It combines the advantages of confocal fluorescence microscopy with fast and precise semi-automatic image analysis. In this work we present how this method was applied to investigate the impact of 310 nm silica nanoparticles on human vascular endothelial cells (HUVEC) in comparison to a cancer cell line derived from the cervix carcinoma (HeLa). The absolute number of intracellular silica nanoparticles within the first 24 h was determined and shown to be cell type-dependent. As a second case study, Particle_in_Cell-3D was used to assess the uptake kinetics of 8 nm and 30 nm ceria nanoparticles interacting with human microvascular endothelial cells (HMEC-1). These small nanoparticles formed agglomerates in biological medium, and the particles that were in effective contact with cells had a mean diameter of 417 nm and 316 nm, respectively. A significant particle size-dependent effect was observed after 48 h of interaction, and the number of intracellular particles was more than four times larger for the 316 nm agglomerates. Interestingly, our results show that for both particle sizes there is a maximum dose of intracellular nanoparticles at about 24 h. One of the causes for such an interesting and unusual uptake behavior could be cell division.

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