Cell Biology

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    Protocols in Current Issue
    Whole-mount Senescence-Associated Beta-Galactosidase (SA-β-GAL) Activity Detection Protocol for Adult Zebrafish
    Authors:  Marta Marzullo, Mounir El Maï and Miguel Godinho Ferreira, date: 07/05/2022, view: 17, Q&A: 0
    [Abstract]

    Senescence-associated beta-galactosidase (SA-β-GAL) is an enzyme that accumulates in the lysosomes of senescent cells, where it hydrolyses β-galactosides. With p16, it represents a well-recognized biomarker used to assess senescence both in vivo and in cell culture. The use of a chromogenic substrate, such as

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    Automated Quantification of Multiple Cell Types in Fluorescently Labeled Whole Mouse Brain Sections Using QuPath
    [Abstract]

    The quantification of labeled cells in tissue sections is crucial to the advancement of biological knowledge. Traditionally, this was a tedious process, requiring hours of careful manual counting in small portions of a larger tissue section. To overcome this, many automated methods for cell analysis have been developed. Recent advances in whole

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    Quantitative Live Confocal Imaging in Aquilegia Floral Meristems
    Authors:  Ya Min, Stephanie J. Conway and Elena M. Kramer, date: 06/20/2022, view: 992, Q&A: 0
    [Abstract]

    In this study, we present a detailed protocol for live imaging and quantitative analysis of floral meristem development in Aquilegia coerulea, a member of the buttercup family (Ranunculaceae). Using confocal microscopy and the image analysis software MorphoGraphX, we were able to examine the cellular growth dynamics during floral organ primordia

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    Labelling of Active Transcription Sites with Argonaute NRDE-3—Image Active Transcription Sites in vivo in Caenorhabditis elegans
    Authors:  Antoine Barrière and Vincent Bertrand, date: 06/05/2022, view: 519, Q&A: 0
    [Abstract]

    Live labelling of active transcription sites is critical to our understanding of transcriptional dynamics. In the most widely used method, RNA sequence MS2 repeats are added to the transcript of interest, on which fluorescently tagged Major Coat Protein binds, and labels transcription sites and transcripts. Here we describe another strategy, using

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    Quantitative Analysis of Actin Cable Length in Yeast
    Authors:  Shane G. McInally, Jane Kondev and Bruce L. Goode, date: 05/05/2022, view: 936, Q&A: 0
    [Abstract]

    Polarized actin cables in S. cerevisiae are linear bundles of crosslinked actin filaments that are assembled by two formins, Bnr1 (localized to the bud neck), and Bni1 (localized to the bud tip). Actin is polymerized at these two sites, which results in cables extending along the cell cortex toward the back of the mother cell. These cables serve

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    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: 1201, 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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    High Throughput Blood-brain Barrier Organoid Generation and Assessment of Receptor-Mediated Antibody Transcytosis
    Authors:  Elena Kassianidou, Claire Simonneau, Alina Gavrilov and Roberto Villaseñor, date: 04/20/2022, view: 668, Q&A: 0
    [Abstract]

    Targeting receptor-mediated transcytosis (RMT) is a successful strategy for drug delivery of biologic agents across the blood-brain barrier (BBB). The recent development of human BBB organoid models is a major advancement to help characterize the mechanisms of RMT and thus accelerate the design of brain delivery technologies. BBB organoids exhibit

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    Single Molecule Tracking Nanoscopy Extended to Two Colors with MTT2col for the Analysis of Cell-Cell Interactions in Leukemia
    Authors:  Loriane Maillot, Magali Irla and Arnauld Sergé, date: 04/20/2022, view: 806, Q&A: 0
    [Abstract]

    Single molecule tracking (SMT) is a powerful technique to study molecular dynamics, and is particularly adapted to monitor the motion and interactions of cell membrane components. Assessing interactions among two molecular populations is classically performed by several approaches, including dual-color videomicroscopy, which allows monitoring of

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    In vivo Ca2+ Imaging in Mouse Salivary Glands
    Authors:  Takahiro Takano and David I. Yule, date: 04/05/2022, view: 781, Q&A: 0
    [Abstract]

    Changes in intracellular calcium drive exocrine cell activity. In the salivary gland, acetylcholine released from parasympathetic neurons mobilizes endoplasmic reticulum calcium stores in acinar cells, which consequently initiates saliva secretion. However, our understanding of the signaling cascade is mainly based on ex vivo studies

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    3D-Structured Illumination Microscopy of Centrosomes in Human Cell Lines
    [Abstract]

    The centrosome is the main microtubule-organizing center of animal cells, and is composed of two barrel-shaped microtubule-based centrioles embedded in protein dense pericentriolar material. Compositional and architectural re-organization of the centrosome drives its duplication, and enables its microtubule-organizing activity and capability to

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