Protocols in Current Issue
    Detection and Quantification of African Swine Fever Virus in MA-104 Cells

    Detection of live African swine fever virus (ASFV) has historically relied on the use of primary swine macrophages (PSM). PSM do not replicate and have to be isolated fresh from donor swine. We previously identified that a MA-104 cells (ATCC #CRL-2378.1), a commercially available cell line isolated from African green monkey (Cercopithecus

    An Imaging Flow Cytometry Method to Measure Citrullination of H4 Histone as a Read-out for Neutrophil Extracellular Traps Formation
    Authors:  Emilia A. Barbu, Venina M. Dominical, Laurel Mendelsohn and Swee Lay Thein, date: 02/20/2021, view: 1115, Q&A: 0

    The formation of neutrophil extracellular traps (NETs) is thought to play a critical role in infections and propagating sterile inflammation. Histone citrullination is an essential and early step in NETs formation, detectable prior to the formation of the hallmark extracellular DNA-scaffolded strands. In addition to the classical microscopy

    Optimized Protocol for the Incorporation of FDAA (HADA Labeling) for in situ Labeling of Peptidoglycan
    Authors:  Katharina Peters, Manuel Pazos, Michael S. VanNieuwenhze and Waldemar Vollmer, date: 08/05/2019, view: 4624, Q&A: 0
    [Abstract] The essential peptidoglycan (PG) layer surrounds the cytoplasmic membrane in nearly all bacteria. It is needed to maintain the shape of the cell and protect it from lysis due to high turgor. Growth of the PG layer is a complex process that involves the activities of PG synthases and hydrolases during elongation and cell division. PG growth sites ...
    Endocytosis Detection in Magnaporthe oryzae
    Authors:  Muxing Liu and Zhengguang Zhang, date: 08/05/2019, view: 2195, Q&A: 0
    [Abstract] Endocytosis is an intracellular trafficking pathway that occurs in nutrient uptake, signal transduction and reconstruction of cell polarity and is conserved in eukaryotic cells. In fungi, endocytosis plays crucial roles in the physiology of hyphal growth and pathogenicity. vidence for endocytosis in filamentous fungi is detected by the ...
    Assessing Membrane Fluidity and Visualizing Fluid Membrane Domains in Bacteria Using Fluorescent Membrane Dyes
    Authors:  Michaela Wenzel, Norbert O. E. Vischer, Henrik Strahl and Leendert W. Hamoen, date: 10/20/2018, view: 6597, Q&A: 0
    [Abstract] Membrane fluidity is a key parameter of bacterial membranes that undergoes quick adaptation in response to environmental challenges and has recently emerged as an important factor in the antibacterial mechanism of membrane-targeting antibiotics. The specific level of membrane fluidity is not uniform across the bacterial cell membrane. Rather, ...
    Detection of Apoptosis-like Cell Death in Ustilago maydis by Annexin V-FITC Staining
    Authors:  Dibya Mukherjee, Aroni Mitra and Anupama Ghosh, date: 08/05/2018, view: 4148, Q&A: 0
    [Abstract] Programmed cell death (PCD) guides the transition between key developmental stages in many organisms. PCD also remains an important fate for many organisms upon exposure to different stress conditions. Therefore, an insight into the progression of PCD during the execution of a biological phenomenon can yield significant details of the underlying ...
    Direct Visualization of the Multicopy Chromosomes in Cyanobacterium Synechococcus elongatus PCC 7942
    Authors:  Ryudo Ohbayashi, Hirofumi Yoshikawa and Satoru Watanabe, date: 08/05/2018, view: 4476, Q&A: 0
    [Abstract] Cyanobacteria are prokaryotic organisms that carry out oxygenic photosynthesis. The fresh water cyanobacterium Synechococcus elongatus PCC 7942 is a model organism for the study of photosynthesis and gene regulation, and for biotechnological applications. Besides several freshwater cyanobacteria, S. elongatus 7942 also contains ...
    Phototaxis Assays of Synechocystis sp. PCC 6803 at Macroscopic and Microscopic Scales
    Authors:  Annik Jakob, Nils Schuergers and Annegret Wilde, date: 06/05/2017, view: 6834, Q&A: 0
    [Abstract] Phototaxis is a mechanism that allows cyanobacteria to respond to fluctuations in the quality and quantity of illumination by moving either towards or away from a light source. Phototactic movement on low concentration agar or agarose plates can be analyzed at macroscopic and microscopic scales representing group behavior and single cell motility, ...
    Fluorescence in situ Localization of Gene Expression Using a lacZ Reporter in the Heterocyst-forming Cyanobacterium Anabaena variabilis
    Authors:  Brenda S. Pratte and Teresa Thiel, date: 01/05/2017, view: 6195, Q&A: 0
    [Abstract] One of the most successful fluorescent proteins, used as a reporter of gene expression in many bacterial, plant and animals, is green fluorescent protein and its modified forms, which also function well in cyanobacteria. However, these fluorescent proteins do not allow rapid and economical quantitation of the reporter gene product, as does the ...
    Quantification of Respiratory Activity in Biofilms
    Authors:  Cláudia N. H. Marques and Scott A. Craver, date: 09/20/2015, view: 8293, Q&A: 1
    [Abstract] Bacteria live mostly as biofilms, not as planktonic cell populations. Bacterial cells living as biofilms are known to be in different physiological status. Persister cells are one of such physiological conditions and they are recognized as to be a stochastically produced sub-population of non-growing bacterial cells. The following protocol ...

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