Cell Biology

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    Protocols in Current Issue
    Detection of Individual RNA in Fixed Cells and Tissues by Chromogenic ISH
    Authors:  Meng Jiang, Chen Lin and Rongqin Ke, date: 02/05/2020, view: 170, Q&A: 0
    [Abstract] Visualization of RNA molecules in situ helps to better understand the functions of expressed genes. Currently, most conventional in situ hybridization methods for visualization of individual RNAs are based on fluorescence detection. Herein we present a chromogenic in situ hybridization protocol for visualization of ...
    Retina Injury and Retina Tissue Preparation to Study Regeneration in Zebrafish
    Authors:  Poonam Sharma and Rajesh Ramachandran, date: 12/20/2019, view: 460, Q&A: 0
    [Abstract] Unlike mammals, primitive vertebrates have immense capability to regenerate almost all of their organs including the central nervous system. Among primitive organisms, zebrafish have been extensively used as a model system for regeneration studies. The retina is a part of the central nervous system and mammals lack the potential to repair any ...
    Analysis of EGF Receptor Endocytosis Using Fluorogen Activating Protein Tagged Receptor
    [Abstract] Functional activities of many transmembrane proteins are controlled by their endocytosis. One of the most studied experimental models is the epidermal growth factor (EGF) receptor (EGFR). However, endocytic trafficking of EGFR has been predominantly analyzed using labeled EGF, whereas quantitative analyses of the endocytosis of the receptor itself ...
    Determination of Chromatin Accessibility in Drosophila Midgut Enterocytes by in situ 5mC Labeling
    Authors:  Eliya Bitman-Lotan, Hector Rincon-Arano, Gal Raz and Amir Orian, date: 11/20/2019, view: 526, Q&A: 0
    [Abstract] Regulation of gene expression involves dynamic changes in chromatin organization, where in many cases open chromatin structure correlates with gene activation. Several methods enable monitoring changes in chromatin accessibility, including ATAC-seq, FAIRE-seq, MNase-seq and DNAse-seq methods, which involve Next-generation-sequencing (NGS). ...
    Leishmania Parasite Quantification by Bioluminescence in Murine Models
    Authors:  Marta Reverte and Nicolas Fasel, date: 11/20/2019, view: 385, Q&A: 0
    [Abstract] Leishmaniasis remains a major public health problem worldwide with a prevalence of 12 million, an incidence of 1 million persons, and 350 million people being at risk. Murine models have been largely used for studying the host-pathogen relationship and developing effective chemotherapies against Leishmania parasites. Thus, preclinical ...
    Ex vivo Drosophila Wing Imaginal Disc Culture and Furin Inhibitor Assay
    Authors:  Alex Sohr, Lijuan Du and Sougata Roy, date: 08/20/2019, view: 838, Q&A: 0
    [Abstract] Furin is an evolutionarily conserved proprotein convertase (PC) family enzyme with a broad range of substrates that are essential for developmental, homeostatic, and disease pathways. Classical genetic approaches and in vitro biochemical or cell biological assays identified that precursor forms of most growth factor family proteins are ...
    A Novel Technique for Imaging and Analysis of Hair Cells in the Organ of Corti Using Modified Sca/eS and Machine Learning
    [Abstract] Here, we describe a sorbitol-based optical clearing method, called modified Sca/eS that can be used to image all hair cells (HCs) in the mouse cochlea. This modification of Sca/eS is defined by three steps: decalcification, de-lipidation, and refractive index matching, which can all be completed within 72 h. Furthermore, we established automated ...
    Characterization of Mouse Adult Testicular Macrophage Populations by Immunofluorescence Imaging and Flow Cytometry
    Authors:  Noushin Mossadegh-Keller and Michael H. Sieweke, date: 03/05/2019, view: 1654, Q&A: 0
    [Abstract] Testicular macrophages (tMΦ) are the most abundant immune cells residing in the testis, an immune-privileged organ. TMΦ are known to exhibit different functions, such as protecting spermatozoa from auto-immune attack by producing immunosuppressive cytokines and trophic roles in supporting spermatogenesis and male sex hormone production. They also ...
    Blinded Visual Scoring of Images Using the Freely-available Software Blinder
    Authors:  Steven D. Cothren, Joel N. Meyer and Jessica H. Hartman, date: 12/05/2018, view: 1958, Q&A: 0
    [Abstract] In nearly all subfields of biomedical sciences, there are phenotypes that are currently classified by expert visual scoring. In research applications, these classifications require the experimenter to be blinded to the treatment group in order to avoid unintentional bias in scoring. Currently, many labs either use laborious and tedious methods to ...
    Staining the Germline in Live Caenorhabditis elegans: Overcoming Challenges by Applying a Fluorescent-dye Feeding Strategy
    Author:  K. Adam Bohnert, date: 11/05/2018, view: 2424, Q&A: 0
    [Abstract] C. elegans provides a tractable model organism for studying germline cell biology. Microscopy experiments are relatively facile, as this worm is transparent and germline development can be observed in real-time using DIC microscopy and/or fluorescent transgenes. Despite these many tools, robust staining techniques for imaging germ cells ...



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