Protocols in Current Issue
    Cryo-transmission Electron Microscopy of Outer-inner Membrane Vesicles Naturally Secreted by Gram-negative Pathogenic Bacteria
    [Abstract] A protocol was developed to visualize and analyze the structure of membrane vesicles (MVs) from Gram-negative bacteria. It is now accepted that these micrometric spherical vesicles are commonly produced by cells from all three domains of life, so the protocol could be useful in the study of vesicles produced by eukaryotes and archaea as well as ...
    Non-invasive Quantification of Cell Wall Porosity by Fluorescence Quenching Microscopy
    Authors:  Xiaohui Liu, Thomas Günther Pomorski and Johannes Liesche, date: 08/20/2019, view: 305, Q&A: 0
    [Abstract] All bacteria, fungi and plant cells are surrounded by a cell wall. This complex network of polysaccharides and glycoproteins provides mechanical support, defines cell shape, controls cell growth and influences the exchange of substances between the cell and its surroundings. Despite its name, the cell wall is a flexible, dynamic structure. ...
    Systematic Quantification of GFP-tagged Protein Foci in Schizosaccharomyces pombe Nuclei
    Authors:  Kim Kiat Lim and Ee Sin Chen, date: 12/20/2018, view: 1434, 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 ...
    Extracting and Integrating Protein Localization Changes from Multiple Image Screens of Yeast Cells
    Authors:  Alex X Lu, Louis-Francois Handfield and Alan M Moses, date: 09/20/2018, view: 1676, Q&A: 0
    [Abstract] The evaluation of protein localization changes in cells under diverse chemical and genetic perturbations is now possible due to the increasing quantity of screens that systematically image thousands of proteins in an organism. Integrating information from different screens provides valuable contextual information about the protein function. For ...
    High-throughput Microscopic Analysis of Salmonella Invasion of Host Cells
    Authors:  Jakub Voznica, Jost Enninga and Virginie Stévenin, date: 09/20/2018, view: 1784, Q&A: 0
    [Abstract] Salmonella is a Gram-negative bacterium causing a gastro-enteric disease called salmonellosis. During the first phase of infection, Salmonella uses its flagella to swim near the surface of the epithelial cells and to target specific site of infection. In order to study the selection criteria that determine which host cells are ...
    A Microfluidic Device for Massively Parallel, Whole-lifespan Imaging of Single Fission Yeast Cells
    Authors:  Stephen K Jones Jr, Eric C Spivey, James R Rybarski and Ilya J Finkelstein, date: 04/05/2018, view: 3290, Q&A: 0
    [Abstract] Whole-lifespan single-cell analysis has greatly increased our understanding of fundamental cellular processes such as cellular aging. To observe individual cells across their entire lifespan, all progeny must be removed from the growth medium, typically via manual microdissection. However, manual microdissection is laborious, low-throughput, and ...
    Live-cell Imaging of Neisseria meningitidis Microcolony Dispersal Induced by Lactate or Other Molecules
    Authors:  Sara Sigurlásdóttir, Olaspers Sara Eriksson, Jens Eriksson and Ann-Beth Jonsson, date: 01/20/2018, view: 2425, Q&A: 0
    [Abstract] To efficiently colonize the nasopharyngeal epithelium, the human restricted pathogen Neisseria meningitidis follows a multistep adhesion cascade. First, the bacteria adhere to host cells and aggregate into spherical shaped structures called microcolonies. Several hours later, single bacteria start dispersing from the microcolonies and ...
    Determination of Local pH Differences within Living Salmonella Cells by High-resolution pH Imaging Based on pH-sensitive GFP Derivative, pHluorin(M153R)
    Authors:  Yusuke V. Morimoto, Nobunori Kami-ike, Keiichi Namba and Tohru Minamino, date: 09/05/2017, view: 3320, Q&A: 0
    [Abstract] The bacterial flagellar type III protein export apparatus is composed of a transmembrane export gate complex and a cytoplasmic ATPase complex. The export apparatus requires ATP hydrolysis and the proton motive force across the cytoplasmic membrane to unfold and transport flagellar component proteins for the construction of the bacterial flagellum ...
    Plasmodium Sporozoite Motility on Flat Substrates
    Authors:  Henriette L Prinz, Julia M Sattler and Friedrich Frischknecht, date: 07/20/2017, view: 4586, Q&A: 0
    [Abstract] Plasmodium sporozoites are the infectious, highly motile forms of the malaria parasite transmitted by Anopheles mosquitoes. Sporozoite motility can be assessed following the dissection of Anopheles salivary glands and isolation of sporozoites in vitro.
    Protein Localization in the Cyanobacterium Anabaena sp. PCC7120 Using Immunofluorescence Labeling
    Authors:  Carla Trigo, Derly Andrade and Mónica Vásquez, date: 06/05/2017, view: 4202, Q&A: 0
    [Abstract] Techniques such as immunoflorescence are widely used to determine subcellular distribution of proteins. Here we report on a method to immunolocalize proteins in Anabaena sp. PCC7120 with fluorophore-conjugated antibodies by fluorescence microscopy. This method improves the permeabilization of cyanobacterial cells and minimizes the ...

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