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Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer

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Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons

Authors: Pavithra Aravamudhan, Krishnan Raghunathan and Terence S. Dermody, date: 11/20/2020, view: 648, Q&A: 0

[Abstract]

Neurotropic reoviruses repurpose host machinery to traffic over long distances in neuronal processes and access distal replication sites. Understanding mechanisms of neuronal transmission is facilitated by using simplified in vitro primary neuronal culture models. Advances in the design of compartmentalized microfluidic devices lend robustness to

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Analysis of Gram-negative Bacteria Peptidoglycan by Ultra-performance Liquid Chromatography

Authors: Laura Alvarez, Baptiste Cordier, Sven van Teeffelen and Felipe Cava, date: 10/05/2020, view: 693, Q&A: 0

[Abstract] Bacteria are surrounded by a protective peptidoglycan cell wall. Provided that this structure and the enzymes involved are the preferred target for our most successful antibiotics, determining its structural and chemical complexity is of the highest interest. Traditionally, high-performance liquid chromatography (HPLC) analyses have been ...

Live-cell Imaging by Super-resolution Confocal Live Imaging Microscopy (SCLIM): Simultaneous Three-color and Four-dimensional Live Cell Imaging with High Space and Time Resolution

Authors: Kazuo Kurokawa and Akihiko Nakano, date: 09/05/2020, view: 1930, Q&A: 0

[Abstract] Many questions in cell biology can be solved by state-of-the-art technology of live cell imaging. One good example is the mechanism of membrane traffic, in which small membrane carriers are rapidly moving around in the cytoplasm to deliver cargo proteins between organelles. For directly visualizing the events in membrane trafficking system, ...

In vitro Cultivation and Visualization of Malaria Liver Stages in Primary Simian Hepatocytes

Authors: Devendra Kumar Gupta and Thierry Diagana, date: 08/20/2020, view: 943, Q&A: 0

[Abstract] Human liver is the primary and obligatory site for malaria infection where sporozoites invade host hepatocytes. Malaria hepatic stages are asymptomatic and represent an attractive target for development of anti-malarial interventions and vaccines. However, owing to lack of robust and reproducible in vitro culture system, it is difficult ...

Live Cell Measurement of the Intracellular pH of Yeast by Flow Cytometry Using a Genetically-Encoded Fluorescent Reporter

Authors: Catherine G. Triandafillou and D. Allan Drummond, date: 06/20/2020, view: 1267, Q&A: 0

[Abstract] The intracellular pH of yeast is a tightly regulated physiological cue that changes in response to growth state and environmental conditions. Fluorescent reporters, which have altered fluorescence in response to local pH changes, can be used to measure intracellular pH. While microscopy is often used to make such measurements, it is relatively ...

Purification of Rice Stripe Virus

Authors: Gang Lu, Min Yao, Yijun Zhou and Xiaorong Tao, date: 03/20/2020, view: 1125, Q&A: 0

[Abstract] Although many spherical and rod-shaped plant virus purification protocols are now available, only a few protocols on filamentous plant virus purification have been published. Here, we report a protocol for large-scale purification of Rice stripe virus (RSV) from RSV-infected rice tissues. RSV virions with high infectivity were first precipitated ...

Rapid Detection of Proliferative Bacteria by Electrical Stimulation

Authors: Conor LA Edwards, Dmitry Malyshev, James P Stratford and Munehiro Asally, date: 02/05/2020, view: 2401, Q&A: 0

[Abstract] Detecting live bacteria is an important task for antimicrobial susceptibility testing (AST) in the medical sector and for quality-monitoring in biological industries. Current methods for live-bacteria detection suffer limitations in speed or sensitivity. In a recent paper, we reported that electrical response dynamics in membrane potential enable ...

Study of Microbial Extracellular Vesicles: Separation by Density Gradients, Protection Assays and Labelling for Live Tracking

Authors: Carolina Coelho, Raghav Vij, Daniel Q. Smith, Nathan R. Brady, Anne Hamacher-Brady and Arturo Casadevall, date: 01/20/2020, view: 1739, Q&A: 0

[Abstract] Extracellular vesicles (EVs) are produced by all domains of life including Bacteria, Archaea and Eukarya. EVs are critical for cellular physiology and contain varied cargo: virulence factors, cell wall remodeling enzymes, extracellular matrix components and even nucleic acids and metabolites. While various protocols for isolating EVs have been ...

A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae

Authors: Abel Cuevas-Bermúdez, Ana I. Garrido-Godino, Francisco Gutiérrez-Santiago and Francisco Navarro, date: 01/05/2020, view: 1575, Q&A: 0

[Abstract] We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using ...

Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves

Authors: Linda Grosche, Katinka Döhner, Alexandra Düthorn, Ana Hickford-Martinez, Alexander Steinkasserer and Beate Sodeik, date: 12/05/2019, view: 4067, Q&A: 0

[Abstract] Given the endemic seroprevalence of herpes simplex viruses (HSV), its associated human diseases, and the emergence of acyclovir-resistant strains, there is a continuous need for better antiviral therapies. Towards this aim, identifying mechanistic details of how HSV-1 manipulates infected cells, how it modulates the immune responses, and how it ...

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