scholarly journals Three-Dimensional Two-Color Dual-Particle Tracking Microscope for Monitoring DNA Conformational Changes and Nanoparticle Landings on Live Cells

ACS Nano ◽  
2020 ◽  
Vol 14 (7) ◽  
pp. 7927-7939
Author(s):  
Yen-Liang Liu ◽  
Evan P. Perillo ◽  
Phyllis Ang ◽  
Mirae Kim ◽  
Duc Trung Nguyen ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Particle Tracking ◽  
Three Dimensional ◽  
Live Cells ◽  
Dna Conformational Changes

scholarly journals Three-dimensional single-particle tracking in live cells: news from the third dimension

2013 ◽  
Vol 15 (7) ◽  
pp. 075008 ◽  
Author(s):  
A Dupont ◽  
M Gorelashvili ◽  
V Schüller ◽  
F Wehnekamp ◽  
D Arcizet ◽  
...  
Keyword(s):  
Particle Tracking ◽  
Single Particle ◽  
Three Dimensional ◽  
Single Particle Tracking ◽  
Live Cells ◽  
The Third ◽  
Third Dimension

Electron Microscopy and image reconstruction reveal the structural basis for spectrin's elastic properties

1992 ◽  
Vol 50 (1) ◽  
pp. 510-511
Author(s):  
Amy M. McGough ◽  
Robert Josephs
Keyword(s):  
Electron Microscopy ◽  
Elastic Properties ◽  
Conformational Changes ◽  
Quaternary Structure ◽  
Three Dimensional ◽  
Membrane Skeleton ◽  
Projection Algorithm ◽  
Structural Basis ◽  
Iterative Approximation ◽  
Membrane Skeletons

The remarkable deformability of the erythrocyte derives in large part from the elastic properties of spectrin, the major component of the membrane skeleton. It is generally accepted that spectrin's elasticity arises from marked conformational changes which include variations in its overall length (1). In this work the structure of spectrin in partially expanded membrane skeletons was studied by electron microscopy to determine the molecular basis for spectrin's elastic properties. Spectrin molecules were analysed with respect to three features: length, conformation, and quaternary structure. The results of these studies lead to a model of how spectrin mediates the elastic deformation of the erythrocyte.Membrane skeletons were isolated from erythrocyte membrane ghosts, negatively stained, and examined by transmission electron microscopy (2). Particle lengths and end-to-end distances were measured from enlarged prints using the computer program MACMEASURE. Spectrin conformation (straightness) was assessed by calculating the particles’ correlation length by iterative approximation (3). Digitised spectrin images were correlation averaged or Fourier filtered to improve their signal-to-noise ratios. Three-dimensional reconstructions were performed using a suite of programs which were based on the filtered back-projection algorithm and executed on a cluster of Microvax 3200 workstations (4).

Adaptive optics enables three-dimensional single particle tracking at the sub-millisecond scale

2013 ◽  
Vol 102 (17) ◽  
pp. 173702 ◽  
Author(s):  
Manuel F. Juette ◽  
Felix E. Rivera-Molina ◽  
Derek K. Toomre ◽  
Joerg Bewersdorf
Keyword(s):  
Adaptive Optics ◽  
Particle Tracking ◽  
Single Particle ◽  
Three Dimensional ◽  
Single Particle Tracking

scholarly journals Parallel, linear, and subnanometric 3D tracking of microparticles with Stereo Darkfield Interferometry

2021 ◽  
Vol 7 (6) ◽  
pp. eabe3902
Author(s):  
Martin Rieu ◽  
Thibault Vieille ◽  
Gaël Radou ◽  
Raphaël Jeanneret ◽  
Nadia Ruiz-Gutierrez ◽  
...  
Keyword(s):  
High Precision ◽  
High Throughput ◽  
Single Molecule ◽  
Particle Tracking ◽  
Focal Plane ◽  
Tracking System ◽  
Three Dimensional ◽  
3D Tracking ◽  
Single Molecule Force Spectroscopy ◽  
A New Technique

While crucial for force spectroscopists and microbiologists, three-dimensional (3D) particle tracking suffers from either poor precision, complex calibration, or the need of expensive hardware, preventing its massive adoption. We introduce a new technique, based on a simple piece of cardboard inserted in the objective focal plane, that enables simple 3D tracking of dilute microparticles while offering subnanometer frame-to-frame precision in all directions. Its linearity alleviates calibration procedures, while the interferometric pattern enhances precision. We illustrate its utility in single-molecule force spectroscopy and single-algae motility analysis. As with any technique based on back focal plane engineering, it may be directly embedded in a commercial objective, providing a means to convert any preexisting optical setup in a 3D tracking system. Thanks to its precision, its simplicity, and its versatility, we envision that the technique has the potential to enhance the spreading of high-precision and high-throughput 3D tracking.

scholarly journals The Core and Holoenzyme Forms of RNA Polymerase from Mycobacterium smegmatis

2018 ◽  
Vol 201 (4) ◽  
Author(s):  
Tomáš Kouba ◽  
Jiří Pospíšil ◽  
Jarmila Hnilicová ◽  
Hana Šanderová ◽  
Ivan Barvík ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Rna Polymerase ◽  
Conformational Changes ◽  
Mycobacterium Smegmatis ◽  
Drug Target ◽  
Conformational Dynamics ◽  
Three Dimensional ◽  
Cryo Electron Microscopy ◽  
Bacterial Rna ◽  
High Degree

ABSTRACT Bacterial RNA polymerase (RNAP) is essential for gene expression and as such is a valid drug target. Hence, it is imperative to know its structure and dynamics. Here, we present two as-yet-unreported forms of Mycobacterium smegmatis RNAP: core and holoenzyme containing σA but no other factors. Each form was detected by cryo-electron microscopy in two major conformations. Comparisons of these structures with known structures of other RNAPs reveal a high degree of conformational flexibility of the mycobacterial enzyme and confirm that region 1.1 of σA is directed into the primary channel of RNAP. Taken together, we describe the conformational changes of unrestrained mycobacterial RNAP. IMPORTANCE We describe here three-dimensional structures of core and holoenzyme forms of mycobacterial RNA polymerase (RNAP) solved by cryo-electron microscopy. These structures fill the thus-far-empty spots in the gallery of the pivotal forms of mycobacterial RNAP and illuminate the extent of conformational dynamics of this enzyme. The presented findings may facilitate future designs of antimycobacterial drugs targeting RNAP.

What controls the unusual melting profiles of small AuNPs/DNA complexes

2019 ◽  
Vol 21 (21) ◽  
pp. 11019-11032 ◽  
Author(s):  
Elia Grueso ◽  
Rosa M. Giráldez-Pérez ◽  
Pilar Perez-Tejeda ◽  
Emilio Roldán ◽  
R. Prado-Gotor
Keyword(s):  
Gold Nanoparticles ◽  
Conformational Changes ◽  
Dna Complexes ◽  
Low Salt ◽  
Dna Conformational Changes

The effect of the addition of low salt concentrations on ds-DNA and ss-DNA conformational changes induced by small N-(2-mercaptopropionyl)glycine gold nanoparticles (AuNPs) is studied in detail by using different techniques. The results are correlated with the unusual melting profiles of the AuNPs/DNA complexes.

scholarly journals Three-Dimensional FRET Reconstruction Microscopy for Analysis of Dynamic Molecular Interactions in Live Cells

2008 ◽  
Vol 95 (1) ◽  
pp. 400-418 ◽  
Author(s):  
Adam D. Hoppe ◽  
Spencer L. Shorte ◽  
Joel A. Swanson ◽  
Rainer Heintzmann
Keyword(s):  
Molecular Interactions ◽  
Three Dimensional ◽  
Live Cells

scholarly journals Three-dimensional particle tracking in microfluidic channel flow using in and out of focus diffraction

2015 ◽  
Vol 45 ◽  
pp. 218-224 ◽  
Author(s):  
Bushra Tasadduq ◽  
Gonghao Wang ◽  
Mohamed El Banani ◽  
Wenbin Mao ◽  
Wilbur Lam ◽  
...  
Keyword(s):  
Channel Flow ◽  
Particle Tracking ◽  
Three Dimensional ◽  
Microfluidic Channel
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