Cell Biology

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    Protocols in Current Issue
    Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence
    Authors:  José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 527, Q&A: 0
    [Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...
    Image-based Quantification of Direct Cell-to-cell Transmission of Bovine Viral Diarrhea Virus
    Authors:  Fernando Merwaiss and Diego E. Alvarez, date: 08/05/2019, view: 396, Q&A: 0
    [Abstract] Different viruses rely on direct cell-to-cell transmission to propagate infection within the infected host. Measuring this mode of transmission in cultured cells is often complicated by the contribution of cell free viruses to spread, and the difficulty to distinguish between primary infected cells that produce the virus and neighboring cells that ...
    QUEEN-based Spatiotemporal ATP Imaging in Budding and Fission Yeast
    Author:  Masak Takaine, date: 08/05/2019, view: 483, Q&A: 0
    [Abstract] Yeasts have provided an exceptional model for studying metabolism and bioenergetics in eukaryotic cells. Among numerous metabolites, adenosine triphosphate (ATP) is a major metabolite that is essential for all living organisms. Therefore, a clearer understanding of ATP dynamics in living yeast cells is important for deciphering cellular energy ...
    Lipid-exchange Rate Assay for Lipid Droplet Fusion in Live Cells
    Authors:  Jia Wang, Boon Tin Chua, Peng Li and Feng-Jung Chen, date: 07/20/2019, view: 684, Q&A: 0
    [Abstract] Lipid droplets (LDs) are central organelles in maintaining lipid homeostasis. Defective LD growth often results in the development of metabolic disorders. LD fusion and growth mediated by cell death–inducing DNA fragmentation factor alpha (DFFA)-like effector (CIDE) family proteins are crucial for various biological processes including unilocular ...
    Hypochlorous Acid Staining with R19-S in the Drosophila Intestine upon Ingestion of Opportunistic Bacteria
    Authors:  Salma Hachfi, Olivia Benguettat and Armel Gallet, date: 05/20/2019, view: 934, Q&A: 0
    [Abstract] The intestine is endowed with an innate immune system that is required to fight any exogenous bacteria that are swallowed along with the food. The first line of defense that is mounted by the gut epithelium is the release of immune Reactive Oxygen Species (ROS), such as hypochlorous acid (HOCl), into the lumen. HOCl is produced within 1.5 h of ...
    Delivering "Chromatic Bacteria" Fluorescent Protein Tags to Proteobacteria Using Conjugation
    Authors:  Rudolf O Schlechter and Mitja NP Remus-Emsermann, date: 04/05/2019, view: 1268, Q&A: 0
    [Abstract] Recently, we published a large and versatile set of plasmids, the chromatic bacteria toolbox, to deliver eight different fluorescent protein genes and four combinations of antibiotic resistance genes to Gram-negative bacteria. Fluorescent tags are important tools for single-cell microbiology, synthetic community studies, biofilm, and host-microbe ...
    On-demand Labeling of SNAP-tagged Viral Protein for Pulse-Chase Imaging, Quench-Pulse-Chase Imaging, and Nanoscopy-based Inspection of Cell Lysates
    Authors:  Roland Remenyi, Raymond Li and Mark Harris, date: 02/20/2019, view: 1657, Q&A: 0
    [Abstract] Advanced labeling technologies allow researchers to study protein turnover inside intact cells and to track the labeled protein in downstream applications. In the context of a viral infection, the combination of imaging and fluorescent labeling of viral proteins sheds light on their biological activity and interaction with the host cell. Initial ...
    Imaging Higher-order Chromatin Structures in Single Cells Using Stochastic Optical Reconstruction Microscopy
    Authors:  Jianquan Xu and Yang Liu, date: 02/05/2019, view: 1796, Q&A: 0
    [Abstract] Higher-order chromatin organization shaped by epigenetic modifications influence the chromatin environment and subsequently regulate gene expression. Direct visualization of the higher-order chromatin structure at their epigenomic states is of great importance for understanding chromatin compaction and its subsequent effect on gene expression and ...
    Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe
    Authors:  Farzane Sivandzade, Aditya Bhalerao and Luca Cucullo, date: 01/05/2019, view: 4203, Q&A: 0
    [Abstract] In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the results across laboratories strongly depends upon following well validated and reliable protocols along with the appropriate controls. Herein, we provide a ...
    Systematic Quantification of GFP-tagged Protein Foci in Schizosaccharomyces pombe Nuclei
    Authors:  Kim Kiat Lim and Ee Sin Chen, date: 12/20/2018, view: 1442, Q&A: 0
    [Abstract] DNA damage repair proteins form foci in response to DNA damaging agents. The efficiency and integrity of the DNA repair pathway of a particular eukaryotic (mutant) strain is usually determined by the number of foci formed compared with their wild-type counterpart. Conventionally, focus number is determined visually, and this low accuracy may ...



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