Cell-based analysis —Microbial cell biology —Microbiology

Real-Time Analysis of Mitochondrial Electron Transport Chain Function in Toxoplasma gondii Parasites Using a Seahorse XFe96 Extracellular Flux Analyzer

Authors: Jenni A. Hayward, Esther Rajendran, F. Victor Makota, Brad J. Bassett, Michael Devoy, Teresa Neeman and Giel G. van Dooren, date: 01/05/2022, view: 909, Q&A: 0

[Abstract]

The mitochondrial electron transport chain (ETC) performs several critical biological functions, including maintaining mitochondrial membrane potential, serving as an electron sink for important metabolic pathways, and contributing to the generation of ATP via oxidative phosphorylation. The ETC is important for the survival of many eukaryotic

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Single Cell Analysis and Sorting of Aspergillus fumigatus by Flow Cytometry

Authors: Gareth Howell and Robert-Jan Bleichrodt, date: 04/20/2021, view: 2617, Q&A: 0

[Abstract]

Experimental results in fungal biology research are usually obtained as average measurements across whole populations of cells, whilst ignoring what is happening at the single cell level. Microscopy has allowed us to study single-cell behavior, but it has low throughput and cannot be used to select individual cells for downstream experiments. Here

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Generation and Implementation of Reporter BHK-21 Cells for Live Imaging of Flavivirus Infection

Authors: Jorge L. Arias-Arias and Rodrigo Mora-Rodríguez, date: 03/05/2021, view: 1917, Q&A: 0

[Abstract]

The genus Flavivirus within the family Flaviviridae includes many viral species of medical importance, such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV), among others. Presently, the identification of flavivirus-infected cells is based on either the immunolabeling of viral proteins, the application of recombinant

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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: 2620, 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 ...

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: 2821, 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: 2765, 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 ...

TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4⁺ T Cells

Authors: Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 5767, Q&A: 1

[Abstract] The latent HIV-1 viral reservoir in resting CD4⁺ (rCD4⁺) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...

Analysis of Early Phase HIV-1 Replication and Integration Events by Using Real-time PCR

Authors: Graf Laura, Steven P. Moran, Sally Ro, Carlos M.C. de Noronha and Binshan Shi, date: 02/20/2019, view: 4234, Q&A: 0

[Abstract] Upon entry into a host cell, the HIV-1 virus undergoes a series of critical early replication events including reverse transcription, nuclear import, and integration of its cDNA into the host genome. Molecular assays used to detect and analyze changes in HIV-1 early phase replication events are valuable tools in developing potential antiretroviral ...

In vivo Analysis of Cyclic di-GMP Cyclase and Phosphodiesterase Activity in Escherichia coli Using a Vc2 Riboswitch-based Assay

Authors: Ying Liu, Hyunhee Kim and Ute Römling, date: 03/05/2018, view: 6018, Q&A: 0

[Abstract] Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that regulates distinct aspects of bacterial physiology. It is synthesized by diguanylate cyclases (DGCs) and hydrolyzed by phosphodiesterases (PDEs). To date, the activities of DGC and PDE are commonly assessed by phenotypic assays, mass spectrometry analysis of ...

Isolation, BODIPY Labeling and Uptake of Exosomes in Hepatic Stellate Cells

Author: Shubham Shrivastava, date: 12/05/2017, view: 9771, Q&A: 1

[Abstract] Exosomes have emerged as an important mediator of intercellular communication. They are present in extracellular milieu and therefore, easily accessible by neighboring or distant cells. They carry mRNA, microRNAs and proteins within their vesicles and once internalized by recipient cells; they can modulate multiple signaling pathways with ...

Microbiology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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