Direct quantitative screening of influenza A virus without DNA amplification by single-particle dual-mode total internal reflection scattering

2017 ◽  
Vol 87 ◽  
pp. 842-849 ◽  
Author(s):  
Seungah Lee ◽  
Suresh Kumar Chakkarapani ◽  
Edward S. Yeung ◽  
Seong Ho Kang
Keyword(s):  
Influenza A Virus ◽  
Single Particle ◽  
Total Internal Reflection ◽  
Influenza A ◽  
Dna Amplification ◽  
Internal Reflection ◽  
Dual Mode ◽  
Quantitative Screening

scholarly journals Functional Characterization of an Electromagnetic Perceptive Protein

2020 ◽  
Author(s):  
Sunayana Mitra ◽  
Carlo Barnaba ◽  
Jens Schmidt ◽  
Galit Pelled ◽  
Assaf A. Gilad
Keyword(s):  
Particle Tracking ◽  
Single Particle ◽  
Total Internal Reflection ◽  
Magnetic Stimulation ◽  
New Technologies ◽  
Cellular Function ◽  
Single Particle Tracking ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Diffusion Dynamics

AbstractMagnetoreception, the response to geomagnetic fields is a well described phenomenon in nature. However, it is likely that convergent evolution led to different mechanisms in different organisms. One intriguing example is the unique Electromagnetic Perceptive Gene (EPG) from the glass catfish Kryptopterus vitreolus, that can remotely control cellular function, upon magnetic stimulation in in-vitro and in animal models. Here, we report for the first time the cellular location and orientation of the EPG protein. We utilized a differential labelling technique in determining that the EPG protein is a membrane anchored protein with an N-terminal extracellular domain. The kinetics and diffusion dynamics of the EPG protein in response to magnetic stimulation was also elucidated using single particle imaging and tracking. Pulse chase labelling and Total Internal Reflection Fluorescence (TIRF) imaging revealed an increase in EPG kinetics post magnetic stimulation activation at a single particle level. Trajectory analysis show notably different EPG protein kinetics before and after magnetic stimulation in both 2 (free vs bound particle) and 3 state (free vs intermediate vs bound particle) tracking models. These data serve to provide additional information that support and understand the underlying biophysical mechanisms behind EPG activation by magnetic stimulation. In conclusion, our results provide evidence for the basis of magnetoreception in EPG protein that would aid in future studies that seek to understand this novel mechanism. This study is important for understanding the phenomenon of magnetoreception as well as developing new technologies for magnetogenetics – the utilization of electromagnetic fields to remotely control cellular function.Graphical TOCElucidation of magnetoreception in a fish derived Electromagnetic Perceptive Gene (EPG), using genetic tagging and single particle tracking with Total Internal Reflection Fluorescence (TIRF) suggests changes in kinetics of membranal motion upon stimulation by magnetic field.

Single-particle spectroscopy and defocused imaging of anisotropic gold nanorods by total internal reflection scattering microscopy

The Analyst ◽  
2020 ◽  
Vol 145 (18) ◽  
pp. 6038-6044 ◽  
Author(s):  
Jaeran Lee ◽  
Geun Wan Kim ◽  
Ji Won Ha
Keyword(s):  
Gold Nanorods ◽  
Single Particle ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Single Particle Spectroscopy ◽  
Defocused Imaging

Total internal reflection scattering (TIRS) microscopy provided higher S/N ratio and more accurate angular information of gold nanorods.

scholarly journals Single-particle evanescent light scattering simulations for total internal reflection microscopy

2006 ◽  
Vol 45 (28) ◽  
pp. 7299 ◽  
Author(s):  
Laurent Helden ◽  
Elena Eremina ◽  
Norbert Riefler ◽  
Christopher Hertlein ◽  
Clemens Bechinger ◽  
...  
Keyword(s):  
Light Scattering ◽  
Single Particle ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Total Internal Reflection Microscopy ◽  
Evanescent Light

scholarly journals Label-Free Single-Particle Imaging of the Influenza Virus by Objective-Type Total Internal Reflection Dark-Field Microscopy

PLoS ONE ◽  
2012 ◽  
Vol 7 (11) ◽  
pp. e49208 ◽  
Author(s):  
Sawako Enoki ◽  
Ryota Iino ◽  
Nobuhiro Morone ◽  
Kunihiro Kaihatsu ◽  
Shouichi Sakakihara ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Single Particle ◽  
Total Internal Reflection ◽  
Dark Field ◽  
Internal Reflection ◽  
Label Free ◽  
Dark Field Microscopy ◽  
Particle Imaging ◽  
Single Particle Imaging ◽  
Objective Type

scholarly journals Target membrane cholesterol modulates single influenza virus membrane fusion efficiency but not rate

2019 ◽  
Author(s):  
K. N. Liu ◽  
S. G. Boxer
Keyword(s):  
Membrane Fusion ◽  
Influenza A Virus ◽  
Receptor Binding ◽  
Single Particle ◽  
Influenza A ◽  
Mixing Efficiency ◽  
Membrane Cholesterol ◽  
Host Membrane ◽  
Target Membrane ◽  
Viral Receptors

AbstractHost lipid composition influences many stages of the influenza A virus (IAV) entry process, including: initial binding of IAV to sialylated glycans, fusion between the viral envelope and the host membrane, and the formation of a fusion pore through which the viral genome is transferred into a target cell. In particular, target membrane cholesterol has been shown to preferentially associate with virus receptors and alter physical properties of the membrane like fluidity and curvature. These properties affect both IAV binding and fusion, which makes it difficult to isolate the role of cholesterol in IAV fusion from receptor binding effects. Here, we develop a new fusion assay that uses synthetic DNA-lipid conjugates as surrogate viral receptors to tether virions to target vesicles. To avoid the possibly perturbative effect of adding a self-quenched concentration of dye-labeled lipids to the viral membrane, we tether virions to lipid-labeled target vesicles, and use fluorescence microscopy to detect individual, pH-triggered IAV membrane fusion events. Through this approach, we find that cholesterol in the target membrane enhances the efficiency of single-particle IAV lipid mixing, while the rate of lipid mixing is independent of cholesterol composition. We also find that the single-particle kinetics of influenza lipid mixing to target membranes with different cholesterol compositions is independent of receptor binding, suggesting that cholesterol-mediated spatial clustering of viral receptors within the target membrane does not significantly affect IAV hemifusion. These results are consistent with the hypothesis that target membrane cholesterol increases lipid mixing efficiency by altering host membrane curvature.Statement of SignificanceInfluenza A virus is responsible for millions of cases of flu each year. In order to replicate, influenza must enter a host cell through virus membrane fusion, and cholesterol in the target membrane is vital to the dynamics of this process. We report a receptor-free, single virus fusion assay that requires no fluorescent labeling of virus particles. We use this assay to show that cholesterol increases the fraction of fusion events in a manner that is correlated with the spontaneous curvature of the target membrane but is independent of receptor binding. This assay represents a promising strategy for studying viral fusion processes of other enveloped viruses.

Antiviral effect of polyphenol-enriched extract of Rumex Acetosa L. against Influenza A virus

Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
A Derksen ◽  
W Hafezi ◽  
A Hensel ◽  
J Kühn
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Antiviral Effect ◽  
Rumex Acetosa
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