Cell Biology

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    Protocols in Current Issue
    Visualization and Purification of Caenorhabditis elegans Germ Granule Proteins Using Proximity Labeling
    Authors:  Hannah L. Hertz, Ian F. Price and Wen Tang, date: 04/20/2022, view: 1056, Q&A: 0
    [Abstract]

    Membraneless organelles, such as germ granules and stress granules, are liquid-like condensates formed by phase transition. Recently, we and others have adopted proximity-based labeling methods to determine the composition of these membraneless compartments. Here, we describe the use of TurboID—an engineered promiscuous biotin

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    Combination of Immunofluorescence and Quantitative Fluorescence In-situ Hybridization for Analysing Differential Gene Expression in the Niche Cells of the Drosophila Lymph Gland
    Authors:  Parvathy Ramesh, Sushmit Ghosh and Lolitika Mandal, date: 01/20/2022, view: 1212, Q&A: 0
    [Abstract]

    The Drosophila larval haematopoietic organ or lymph gland consists of multiple cell types arranged in zones. The smallest stem cell compartment consists of 40-45 cells that constitute the haematopoietic niche. In order to analyse the haematopoietic niche, it needs to be labelled with a specific antibody to differentiate it from the other cell

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    Non-invasive Imaging of Rice Roots in Non-compacted and Compacted Soil
    Authors:  Bipin K. Pandey, Jonathan A. Atkinson and Craig J. Sturrock, date: 12/20/2021, view: 1778, Q&A: 0
    [Abstract]

    Roots are the prime organ for nutrient and water uptake and are therefore fundamental to the growth and development of plants. However, physical challenges of a heterogeneous environment and diverse edaphic stresses affect root growth in soil. Compacted soil is a serious global problem, causing inhibition of root elongation, which reduces surface

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    Pericyte Mapping in Cerebral Slices with the Far-red Fluorophore TO-PRO-3
    [Abstract]

    This protocol describes a method for high-resolution confocal imaging of pericytes with the far-red fluorophore TO-PROTM-3 Iodide 642/661 in cerebral slices of murine. Identification of pericytes with TO-PRO-3 is a short time-consuming, high cost-effective and robust technique to label pericytes with no need for immunostaining or generation of

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    Collection of in vivo Capacitated Sperm from Different Locations Along the Reproductive Tract of Time-Mated Female Mice by Microdissection
    Authors:  Lukas Ded and Jean-Ju Chung, date: 10/20/2021, view: 1341, Q&A: 0
    [Abstract]

    Mammalian sperm cells are not capable of fertilizing an egg immediately after ejaculation; instead, they must gradually acquire the capacity to fertilize while they travel inside the female reproductive tract. Sperm cells are transported by the muscular activity of the myometrium to the utero-tubal junction (UTJ) before entering the oviduct where

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    Microscopic Detection of ASC Inflammasomes in Bone Marrow Derived Macrophages Post Stimulation
    Authors:  Longjun Li, Rudi Mao and Xiaopeng Qi, date: 09/05/2021, view: 1684, Q&A: 0
    [Abstract]

    An inflammasome is an intracellular multiprotein complex that plays important roles in host defense and inflammatory responses. Inflammasomes are typically composed of the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), cytoplasmic sensor protein, and the effector protein pro-caspase-1. ASC assembly into a protein

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    Fluorescence-based Single-cell Analysis of Whole-mount-stained and Cleared Microtissues and Organoids for High Throughput Screening
    Authors:  Henrik Renner, Mandy Otto, Martha Grabos, Hans R. Schöler and Jan M. Bruder, date: 06/20/2021, view: 2818, Q&A: 1
    [Abstract]

    Three-dimensional (3D) cell culture, especially in the form of organ-like microtissues (“organoids”), has emerged as a novel tool potentially mimicking human tissue biology more closely than standard two-dimensional culture. Typically, tissue sectioning is the standard method for immunohistochemical analysis. However, it removes cells

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    Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium
    Authors:  Leonie Witte, Karen Linnemannstöns, Mona Honemann-Capito and Julia Christina Gross, date: 06/05/2021, view: 2216, Q&A: 0
    [Abstract]

    Secretory Wnt trafficking can be studied in the polarized epithelial monolayer of Drosophila wing imaginal discs (WID). In this tissue, Wg (Drosophila Wnt-I) is presented on the apical surface of its source cells before being internalized into the endosomal pathway. Long-range Wg secretion and spread depend on secondary secretion from endosomal

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    Retention Using Selective Hooks (RUSH) Cargo Sorting Assay for Protein Vesicle Tracking in HeLa Cells
    Authors:  Natalia Pacheco-Fernandez, Mehrshad Pakdel and Julia von Blume, date: 03/05/2021, view: 3809, Q&A: 0
    [Abstract]

    Monitoring vesicle trafficking is an excellent tool for the evaluation of protein dynamics in living cells. Such study is key for the understanding of protein sorting and secretion. Recent developments in microscopy, as well as new methodologies developed to study synchronized trafficking of proteins, allowed a better understanding of signaling,

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    Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva
    Authors:  Raffaella Klima, Giulia Romano, Monsurat Gbadamosi, Aram Megighian and Fabian Feiguin, date: 02/05/2021, view: 1758, Q&A: 0
    [Abstract]

    Alterations in synaptic transmission are critical early events in neuromuscular disorders. However, reliable methodologies to analyze the functional organization of the neuromuscular synapses are still needed. This manuscript provides a detailed protocol to analyze the molecular assembly of the neuromuscular synapses through

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