Cell Biology

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    Protocols in Current Issue
    Colocalization Analysis for Cryosectioned and Immunostained Tissue Samples with or without Label Retention Expansion Microscopy (LR-ExM) by JACoP
    Authors:  Xiang Zhao, Jason Garcia, Loic A. Royer and Su Guo, date: 03/05/2022, view: 1763, Q&A: 0
    [Abstract]

    Asymmetric cell division (ACD) is fundamental for balancing cell proliferation and differentiation in metazoans. During active neurogenesis in the developing zebrafish forebrain, radial glia progenitors (RGPs) mainly undergo ACD to produce one daughter with high activity of Delta/Notch signaling (proliferative cell fate) and another daughter with

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    Labeling Endogenous Proteins Using CRISPR-mediated Insertion of Exon (CRISPIE)
    Authors:  Evan A. Wilson, Tianyi Mao and Haining Zhong, date: 03/05/2022, view: 1876, Q&A: 0
    [Abstract]

    The CRISPR/Cas9 technology has transformed our ability to edit eukaryotic genomes. Despite this breakthrough, it remains challenging to precisely knock-in large DNA sequences, such as those encoding a fluorescent protein, for labeling or modifying a target protein in post-mitotic cells. Previous efforts focusing on sequence insertion to the

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    Labeling and Tracking Mitochondria with Photoactivation in Drosophila Embryos
    Authors:  Sayali Chowdhary and Richa Rikhy, date: 03/05/2022, view: 1270, Q&A: 0
    [Abstract]

    Mitochondria are relatively small, fragmented, and abundant in the large embryos of Drosophila, Xenopus and zebrafish. It is essential to study their distribution and dynamics in these embryos to understand the mechanistic role of mitochondrial function in early morphogenesis events. Photoactivation of mitochondrially tagged GFP (mito-PA-GFP) is

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    Quantitative Determination of Primary Cilia Protein Distribution Using Immunofluorescence Staining and MATLAB Analysis
    Authors:  Andrew D. Jenks, Marc Fivaz and Barbara E. Tanos, date: 12/05/2021, view: 1779, Q&A: 1
    [Abstract]

    Primary cilia are microtubule-based sensory organelles surrounded by membrane. They can detect mechanical and chemical stimuli. The last few years have uncovered cilia as unique signaling hubs that host a number of receptors and effector molecules. Thus, defining how specific proteins localize and are distributed along the cilium is critical to

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    Whole-mount Staining of Mouse Diaphragm Neuromuscular Junctions
    Authors:  Rula Sha, Zhenzhen Wang, Xue You, Yuguo Liu, Zhiqin Xie and Ying Feng, date: 11/05/2021, view: 1087, Q&A: 0
    [Abstract]

    The neuromuscular junction (NMJ) is a specialized synapse that connects the terminal end of a motor neuron and a skeletal muscle fiber. Defects in NMJ cause abnormalities of neuromuscular transmission, leading to NMJ disorders. The mammalian diaphragm muscle is essential for respiration and has been widely used to study NMJ formation. Here, we

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    Fixation and Immunostaining of Endogenous Proteins or Post-translational Modifications in Caenorhabditis elegans
    Authors:  Robert O'Hagan and Irini Topalidou, date: 10/05/2021, view: 1610, Q&A: 0
    [Abstract]

    Although the advent of genetically-encoded fluorescent markers, such as the green fluorescent protein (GFP; Chalfie et al., 1994), has enabled convenient visualization of gene expression in vivo, this method is generally not effective for detecting post-translational modifications because they are not translated from DNA sequences.

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    Spatiotemporal Quantification of Cytosolic pH in Arabidopsis Pollen Tubes
    Authors:  Maria Teresa Portes, Daniel S.C. Damineli and José A. Feijó, date: 07/20/2021, view: 1515, Q&A: 0
    [Abstract]

    Ion-specific probes and fluorescent indicators have been key in establishing the role of ion signaling, namely calcium, protons, and anions, in plant development, providing a robust approach for monitoring spatiotemporal changes in intracellular ion dynamics. The integration of protons/pH in signaling mechanisms is especially important as reports

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    A Multi-color Bicistronic Biosensor to Compare the Translation Dynamics of Different Open Reading Frames at Single-molecule Resolution in Live Cells
    Authors:  Amanda L. Koch, Tatsuya Morisaki and Timothy J. Stasevich, date: 07/20/2021, view: 1788, Q&A: 0
    [Abstract]

    Here, we describe how to image and quantitate the translation dynamics of a bicistronic biosensor that we recently created to fairly compare cap-dependent and IRES-mediated translation at single-molecule resolution in live human cells. This technique employs a pair of complementary intrabodies loaded into living cells that co-translationally bind

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    Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content
    Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 2227, Q&A: 0
    [Abstract]

    Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several aspects of C. elegans sperm quality, including number, size, rate of activation, and mitochondrial

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    In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
    [Abstract]

    Adenosine 5’-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms

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