Mammalia

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    Protocols in Current Issue
    Evaluation of Genotoxicity by Micronucleus Assay in vitro and by Allium cepa Test in vivo
    Authors:  Christina N. Banti and Sotiris K. Hadjikakou, date: 07/20/2019, view: 758, Q&A: 0
    [Abstract] The in vitro and in vivo genotoxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) is evaluated by the micronucleus test and the Allium cepa assay, respectively. Fetal lung fibroblast cells (MRC-5), normal human corneal epithelial cells (HCEC) and ...
    Physical Removal of the Midbody Remnant from Polarised Epithelial Cells Using Take-Up by Suction Pressure (TUSP)
    Authors:  Miguel Bernabé-Rubio, David C. Gershlick and Miguel A. Alonso, date: 04/20/2017, view: 4646, Q&A: 0
    [Abstract] In polarised epithelial cells the midbody forms at the apical cell surface during cytokinesis. Once severed, the midbody is inherited by one of the daughter cells remaining tethered to the apical plasma membrane where it participates in non-cytokinetic processes, such as primary ciliogenesis. Here, we describe a novel method to physically remove ...
    Bone Resorption Assay
    Authors:  Carina Scholtysek, Gerhard Krönke and Georg Schett, date: 07/20/2014, view: 10357, Q&A: 0
    [Abstract] The Bone resorption assay provides an easy to use protocol for quantitatively measuring in vitro osteoclast-mediated bone resorption. Osteoclasts can be seeded onto the bone slices and formation of resorption pits can be quantified via toluidinblue staining (Scholtysek et al., 2013).
    Lysosomal Stability Assay
    Authors:  Nikolaj H. T. Petersen, Thomas Kirkegaard and Marja Jäättelä, date: 06/20/2014, view: 11579, Q&A: 0
    [Abstract] This assay makes use of the dye Acridine Orange (AO) to determine the stability of lysosomes in living cells upon exposure to a confocal microscope laser.

    AO is a lipophilic amine that readily diffuses into cells (Figure 1). Inside the cell it enters the acidic lysosomal compartment where it is protonated and sequestered, shifting its ...
    Flow Cytometric Analysis of Autophagic Activity with Cyto-ID Staining in Primary Cells
    [Abstract] Flow cytometry allows very sensitive and reliable high-throughput analysis of autophagic flux. This methodology permits to screen cells in flow and capture multi-component images. Using this technology autophagic flux may be analysed accurately in both suspension as well as adherent cells upon trypsinization independent of how heterogeneous the ...
    Flow Cytometric Analyses of Autophagic Activity using LC3-GFP fluorescence
    Authors:  Metodi Stankov, Diana Panayotova-Dimitrova, Martin Leverkus and Georg Behrens, date: 04/05/2014, view: 9203, Q&A: 0
    [Abstract] Flow cytometry allows very sensitive and reliable high-throughput analysis of autophagic flux. This methodology permits to screen cells in flow and capture multi-component images. Using this technology autophagic flux may be analysed accurately in both suspension as well as adherent cells upon trypsinization independent of how heterogeneous the ...
    Spindle Angle Measurements
    Authors:  Julien Cau, Nathalie Morin and Guillaume Bompard, date: 10/05/2013, view: 7813, Q&A: 0
    [Abstract] Spindle angles measures derive from the measures of spindle poles positions that were taken from fixed and immunostained adherent cells. To determine spindle angles (α), z-stack images of metaphasic cells immunostained with anti γ-tubulin (spindle poles) and anti β-tubulin antibodies (mitotic spindle) were acquired. A very simple ImageJ software ...
    Transmission Electron Microscopy (TEM) Protocol: Observation Details within Cells
    Authors:  Wei-Haw Peng, Kuo-Shyan Lu, Shu-Mei Lai, Horng-Tzer Shy and Hsiu-Ni Kung, date: 07/05/2013, view: 10515, Q&A: 0
    [Abstract] Transmission electron microscopy is a technique for observing the fine details of organelles in cells or tissues. This protocol is to be used to exam the membrane structure in cells with or without virus infection. Modifications should be made if users want to get images from tissues.



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