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    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: 233, 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 ...
    Ciliary Assembly/Disassembly Assay in Non-transformed Cell Lines
    Authors:  Masaki Saito, Kensuke Sakaji, Wataru Otsu and Ching-Hwa Sung, date: 03/20/2018, view: 450, Q&A: 0
    [Abstract] The primary cilium is a non-motile sensory organelle whose assembly and disassembly are closely associated with cell cycle progression. The primary cilium is elongated from the basal body in quiescent cells and is resorbed as the cells re-enter the cell cycle. Dysregulation of ciliary dynamics has been linked with ciliopathies and other human ...
    Quantitative Live-cell Reporter Assay for Noncanonical Wnt Activity
    Authors:  Edith P. Karuna, Michael W. Susman and Hsin-Yi Henry Ho, date: 03/20/2018, view: 457, Q&A: 0
    [Abstract] Noncanonical Wnt signaling functions independently of the β-catenin pathway to control diverse developmental processes, and dysfunction of the pathway contributes to a number of human pathological conditions, including birth defects and metastatic cancer. Progress in the field, however, has been hampered by the scarcity of functional assays for ...
    Measurement of Intracellular ROS in Caenorhabditis elegans Using 2’,7’-Dichlorodihydrofluorescein Diacetate
    Authors:  Dong Suk Yoon, Myon-Hee Lee and Dong Seok Cha, date: 03/20/2018, view: 440, Q&A: 0
    [Abstract] Reactive oxygen species (ROS) are generated during normal metabolic processes under aerobic conditions. Since ROS production initiates harmful radical chain reactions on cellular macromolecules, including lipid peroxidation, DNA mutation, and protein denaturation, it has been implicated in a wide spectrum of diseases such as cancer, ...
    Construction and Cloning of Minigenes for in vivo Analysis of Potential Splice Mutations
    Authors:  Lisa Maria Riedmayr, Sybille Böhm, Stylianos Michalakis and Elvir Becirovic, date: 03/05/2018, view: 358, Q&A: 0
    [Abstract] Disease-associated mutations influencing mRNA splicing are referred to as splice mutations. The majority of splice mutations are found on exon-intron boundaries defining canonical donor and acceptor splice sites. However, mutations in the coding region (exonic mutations) can also affect mRNA splicing. Exact knowledge of the disease mechanism of ...
    Terminal Deoxynucleotidyl Transferase Mediated Production of Labeled Probes for Single-molecule FISH or RNA Capture
    Authors:  Imre Gaspar, Frank Wippich and Anne Ephrussi, date: 03/05/2018, view: 461, Q&A: 0
    [Abstract] Arrays of short, singly-labeled ssDNA oligonucleotides enable in situ hybridization with single molecule sensitivity and efficient transcript specific RNA capture. Here, we describe a simple, enzymatic protocol that can be carried out using basic laboratory equipment to convert arrays of PCR oligos into smFISH and RAP probesets in a ...
    Sebinger Culture: A System Optimized for Morphological Maturation and Imaging of Cultured Mouse Metanephric Primordia
    Authors:  Mona Elhendawi and Jamie A. Davies, date: 02/20/2018, view: 401, Q&A: 0
    [Abstract] Here, we present a detailed protocol on setting up embryonic renal organ cultures using a culture method that we have optimised for anatomical maturation and imaging. Our culture method places kidney rudiments on glass in a thin film of medium, which results in very flat cultures with all tubules in the same image plane. For reasons not yet ...
    How to Catch a Smurf? – Ageing and Beyond…
    In vivo Assessment of Intestinal Permeability in Multiple Model Organisms
    [Abstract] The Smurf Assay (SA) was initially developed in the model organism Drosophila melanogaster where a dramatic increase of intestinal permeability has been shown to occur during aging (Rera et al., 2011). We have since validated the protocol in multiple other model organisms (Dambroise et al., 2016) and have utilized the ...
    Preparation of Precisely Oriented Cryosections of Undistorted Drosophila Wing Imaginal Discs for High Resolution Confocal Imaging
    Authors:  Samuel Petshow and Marcel Wehrli, date: 02/05/2018, view: 498, Q&A: 0
    [Abstract] The combination of immunofluorescence and laser scanning confocal microscopy (LSM) is essential to high-resolution detection of molecular distribution in biological specimens. A frequent limitation is the need to image deep inside a tissue or in a specific plane, which may be inaccessible due to tissue size or shape. Recreating high-resolution 3D ...
    Ex vivo Trophoblast-specific Genetic Manipulation Using Lentiviral Delivery
    Authors:  Damayanti Chakraborty, Masanaga Muto and Michael J. Soares, date: 12/20/2017, view: 675, Q&A: 0
    [Abstract] In this protocol report, we describe a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Lentiviral packaged gene constructs can be efficiently and specifically delivered to the trophoblast cell lineage of the blastocyst. The consequences of ‘gain-of-function’ and ‘loss-of-function’ blastocyst ...