Cell Biology

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    Protocols in Current Issue
    Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts

    With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the

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    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 448, Q&A: 0
    Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the ...
    In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts
    Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long ...
    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: 358, Q&A: 0

    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

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    Serial Cryomicrotomy of Saccharomyces cerevisiae for Serial Electron Cryotomography
    Authors:  Cai Tong Ng, Mark S. Ladinsky and Lu Gan, date: 11/20/2020, view: 407, Q&A: 0
    Electron cryotomography (cryo-ET) is an increasingly popular technique to study cellular structures and macromolecules in situ. Due to poor penetration of electrons through thick biological samples, the vitreously frozen samples for cryo-ET ...
    Serial Cryomicrotomy of Saccharomyces cerevisiae for Serial Electron Cryotomography
    Authors:  Cai Tong Ng, Mark S. Ladinsky and Lu Gan, date: 11/20/2020, view: 407, Q&A: 0
    [Abstract] Electron cryotomography (cryo-ET) is an increasingly popular technique to study cellular structures and macromolecules in situ. Due to poor penetration of electrons through thick biological samples, the vitreously frozen samples for cryo-ET need to be thin. For frozen-hydrated cells, such samples can be produced either by cryomicrotomy or ...
    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: 358, 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

    ...
    In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts
    [Abstract] Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long distance at a high speed in the postnatal mammalian brain, and are thus a powerful model to analyze the ...
    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 448, Q&A: 0
    [Abstract] Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the granuloma’s environment may lead to new therapeutic or preventive tools to improve the control of the ...
    Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts
    [Abstract]

    With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the isolation and characterization of mammary epithelial cell subpopulations have been thoroughly developed for

    ...
    Construction and Validation of A Low-cost, Small-scale, Multiplex Continuous Culturing System for Microorganisms
    Authors:  Lilit Tonoyan, Freddy Guihéneuf, Ruari Friel and Vincent O’Flaherty, date: 11/05/2020, view: 448, Q&A: 0
    [Abstract]

    This protocol details the construction of a simple, low-cost, small-scale, multiplex chemostat system designed for the continuous cultivation of microorganisms in suspension (i.e., bacteria, yeast, microalgae). The continuous culture device can operate at a working volume of 25 ml and allows the run of 8 chemostats in parallel by a single person.

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    Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels
    Authors:  Eric Parigoris, Daniel L. Dunkelmann and Unai Silvan, date: 11/05/2020, view: 770, Q&A: 0
    [Abstract] Giant unilamellar vesicles (GUVs) are a widely used model system for a range of applications including membrane biophysics, drug delivery, and the study of actin dynamics. While several protocols have been developed for their generation in recent years, the use of these techniques involving charged lipid types and buffers of physiological ionic ...
    Super-resolution Imaging of the T cell Central Supramolecular Signaling Cluster Using Stimulated Emission Depletion Microscopy
    Authors:  Sin Lih Tan, Dominic Alibhai, Stephen J. Cross, Harry Thompson and Christoph Wülfing, date: 11/05/2020, view: 425, Q&A: 0
    [Abstract]

    Supramolecular signaling assemblies are of interest for their unique signaling properties. A µm scale signaling assembly, the central supramolecular signaling cluster (cSMAC), forms at the center interface of T cells activated by antigen presenting cells (APC). The adaptor protein linker for activation of T cells (LAT) is a key cSMAC component.

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    K+ Release Assay and K+ Measurement in Oocyte Assay
    Author:  Hong Li, date: 11/05/2020, view: 546, Q&A: 0
    [Abstract] The Xenopus oocyte is a powerful system for the exogenous expression and functional characterization of plant membrane transport proteins. Until now, a number of potassium transporters and channels have been identified in oocytes expression system by the two-electrode voltage clamp technology. It is difficult to characterize K+ ...
    Isolation and CryoTEM of Phages Infecting Bacterial Wine Spoilers
    [Abstract]

    With the objective to isolate phages infecting wine bacterial spoilers, we designed a method for the isolation and purification of phages infecting grape-associated bacteria. The method proved successful to isolate GC1 tectivirus infecting the acetic acid bacterium Gluconobacter cerinus. The isolated phage represents a new genus within the

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