Membrane Protein Clusters at Nanoscale Resolution: More Than Pretty Pictures

Physiology ◽  
2010 ◽  
Vol 25 (2) ◽  
pp. 116-124 ◽  
Author(s):  
Thorsten Lang ◽  
Silvio O. Rizzoli
Keyword(s):  
Membrane Protein ◽  
Fluorescence Microscopy ◽  
Molecular Machines ◽  
Protein Conformations ◽  
Superresolution Microscopy ◽  
Protein Clusters ◽  
Microscopy Techniques

Fluorescence microscopy is powerful for analyzing the composition and dynamics of cellular elements, but studying precise molecule patterns is precluded due to diffraction limited resolution. This barrier has been lifted now through several superresolution microscopy techniques. They revealed that proteins assemble in defined groups (clusters). A new challenge thus appears for the biologist: to find out whether clusters are molecular machines, stabilizers of defined protein conformations, or simply protein reservoirs.

scholarly journals Transcranial in vivo detection of amyloid-beta at single plaque resolution with large-field multifocal illumination fluorescence microscopy

2020 ◽  
Author(s):  
Ruiqing Ni ◽  
Zhenyue Chen ◽  
Gloria Shi ◽  
Alessia Villois ◽  
Quanyu Zhou ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Fluorescence Microscopy ◽  
Amyloid Beta ◽  
Large Field ◽  
Microscopy Technique ◽  
In Vivo Detection ◽  
Amyloid Deposits ◽  
Microscopy Techniques

AbstractThe abnormal deposition of beta-amyloid proteins in the brain is one of the major histopathological hallmarks of Alzheimer’s disease. Currently available intravital microscopy techniques for high-resolution plaque visualization commonly involve highly invasive procedures and are limited to a small field-of-view within the rodent brain. Here, we report the transcranial detection of amyloid-beta deposits at the whole brain scale with 20 μm resolution in APP/PS1 and arcAβ mouse models of Alzheimer’s disease amyloidosis using a large-field multifocal (LMI) fluorescence microscopy technique. Highly sensitive and specific detection of amyloid-beta deposits at a single plaque level in APP/PS1 and arcAβ mice was facilitated using luminescent conjugated oligothiophene HS-169. Immunohistochemical staining with HS-169, anti-Aβ antibody 6E10, and conformation antibodies OC (fibrillar) of brain tissue sections further showed that HS-169 resolved compact parenchymal and vessel-associated amyloid deposits. The novel imaging platform offers new prospects for in vivo studies into Alzheimer’s disease mechanisms in animal models as well as longitudinal monitoring of therapeutic responses at a single plaque level.

Heterogeneity of milk fat globule membrane structure and composition as observed using fluorescence microscopy techniques

2008 ◽  
Vol 18 (12) ◽  
pp. 1081-1089 ◽  
Author(s):  
Jaap M. Evers ◽  
Richard G. Haverkamp ◽  
Stephen E. Holroyd ◽  
Geoffrey B. Jameson ◽  
Duncan D.S. Mackenzie ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Milk Fat ◽  
Membrane Structure ◽  
Milk Fat Globule Membrane ◽  
Milk Fat Globule ◽  
Fat Globule ◽  
Microscopy Techniques

scholarly journals Visualization of mt nucleoids by superresolution microscopy techniques

2012 ◽  
Vol 1817 ◽  
pp. S154-S155
Author(s):  
Andrea Dlasková ◽  
Mike Mlodzianoski ◽  
Tomáš Špacek ◽  
Jitka Šantorová ◽  
Jan Tauber ◽  
...  
Keyword(s):  
Superresolution Microscopy ◽  
Microscopy Techniques

scholarly journals Characterization of Mesenchymal Stem Cell Differentiation within Miniaturized 3D Scaffolds through Advanced Microscopy Techniques

2020 ◽  
Vol 21 (22) ◽  
pp. 8498
Author(s):  
Valentina Parodi ◽  
Emanuela Jacchetti ◽  
Arianna Bresci ◽  
Benedetta Talone ◽  
Carlo M. Valensise ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Fluorescence Microscopy ◽  
Fluorescent Dyes ◽  
Phenotypic Expression ◽  
Stem Cell Differentiation ◽  
Confocal Fluorescence Microscopy ◽  
Force Transmission ◽  
Confocal Fluorescence ◽  
Microscopy Techniques

Three-dimensional culture systems and suitable substrates topographies demonstrated to drive stem cell fate in vitro by mechanical conditioning. For example, the Nichoid 3D scaffold remodels stem cells and shapes nuclei, thus promoting stem cell expansion and stemness maintenance. However, the mechanisms involved in force transmission and in biochemical signaling at the basis of fate determination are not yet clear. Among the available investigation systems, confocal fluorescence microscopy using fluorescent dyes enables the observation of cell function and shape at the subcellular scale in vital and fixed conditions. Contrarily, nonlinear optical microscopy techniques, which exploit multi-photon processes, allow to study cell behavior in vital and unlabeled conditions. We apply confocal fluorescence microscopy, coherent anti-Stokes Raman scattering (CARS), and second harmonic generation (SHG) microscopy to characterize the phenotypic expression of mesenchymal stem cells (MSCs) towards adipogenic and chondrogenic differentiation inside Nichoid scaffolds, in terms of nuclear morphology and specific phenotypic products, by comparing these techniques. We demonstrate that the Nichoid maintains a rounded nuclei during expansion and differentiation, promoting MSCs adipogenic differentiation while inhibiting chondrogenesis. We show that CARS and SHG techniques are suitable for specific estimation of the lipid and collagenous content, thus overcoming the limitations of using unspecific fluorescent probes.

Imaging the invisible: resolving cellular microcompartments by superresolution microscopy techniques

2013 ◽  
Vol 394 (9) ◽  
pp. 1097-1113 ◽  
Author(s):  
Michael Hensel ◽  
Jürgen Klingauf ◽  
Jacob Piehler
Keyword(s):  
Imaging Techniques ◽  
Live Cell ◽  
Temporal Organization ◽  
Membrane Microdomains ◽  
Superresolution Microscopy ◽  
Neuronal Synapses ◽  
Spatio Temporal ◽  
Microscopy Techniques ◽  
Plasma Membrane Microdomains ◽  
Diffraction Barrier

Abstract Unraveling the spatio-temporal organization of dynamic cellular microcompartments requires live cell imaging techniques capable of resolving submicroscopic structures. While the resolution of traditional far-field fluorescence imaging techniques is limited by the diffraction barrier, several fluorescence-based microscopy techniques providing sub-100 nm resolution have become available during the past decade. Here, we briefly introduce the optical principles of these techniques and compare their capabilities and limitations with respect to spatial and temporal resolution as well as live cell capabilities. Moreover, we summarize how these techniques contributed to a better understanding of plasma membrane microdomains, the dynamic nanoscale organization of neuronal synapses and the sub-compartmentation of microorganisms. Based on these applications, we highlight complementarity of these techniques and their potential to address specific challenges in the context of dynamic cellular microcompartments, as well as the perspectives to overcome current limitations of these methods.

scholarly journals Comparing phototoxicity during the development of a zebrafish craniofacial bone using confocal and light sheet fluorescence microscopy techniques

2012 ◽  
Vol 6 (11-12) ◽  
pp. 920-928 ◽  
Author(s):  
Matthew Jemielita ◽  
Michael J. Taormina ◽  
April DeLaurier ◽  
Charles B. Kimmel ◽  
Raghuveer Parthasarathy
Keyword(s):  
Fluorescence Microscopy ◽  
Light Sheet ◽  
Craniofacial Bone ◽  
Microscopy Techniques ◽  
Light Sheet Fluorescence Microscopy

scholarly journals Deciphering tissue morphodynamics using bioimage informatics

2017 ◽  
Vol 372 (1720) ◽  
pp. 20150512 ◽  
Author(s):  
Alexandre C. Dufour ◽  
Anneliene H. Jonker ◽  
Jean-Christophe Olivo-Marin
Keyword(s):  
Developmental Biology ◽  
Image Analysis ◽  
Fluorescence Microscopy ◽  
Automated Analysis ◽  
Tissue Level ◽  
Bioimage Informatics ◽  
Software Packages ◽  
Introductory Overview ◽  
To Come ◽  
Microscopy Techniques

In recent years developmental biology has greatly benefited from the latest advances in fluorescence microscopy techniques. Consequently, quantitative and automated analysis of this data is becoming a vital first step in the quest for novel insights into the various aspects of development. Here we present an introductory overview of the various image analysis methods proposed for developmental biology images, with particular attention to openly available software packages. These tools, as well as others to come, are rapidly paving the way towards standardized and reproducible bioimaging studies at the whole-tissue level. Reflecting on these achievements, we discuss the remaining challenges and the future endeavours lying ahead in the post–image analysis era. This article is part of the themed issue ‘Systems morphodynamics: understanding the development of tissue hardware’.

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