Cell Biology

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    Protocols in Current Issue
    A Method for Estimating the Potential Synaptic Connections Between Axons and Dendrites From 2D Neuronal Images
    Authors:  Carolina Tecuatl, Diek W. Wheeler and Giorgio A. Ascoli, date: 07/05/2021, view: 1283, Q&A: 0
    [Abstract]

    Computational neuroscience aims to model, reproduce, and predict network dynamics for different neuronal ensembles by distilling knowledge derived from electrophysiological and morphological evidence. However, analyses and simulations often remain critically limited by the sparsity of direct experimental constraints on essential parameters, such

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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: 1097, 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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    Whole-mount Immunohistochemistry of Adult Zebrafish Retina for Advanced Imaging
    Authors:  Mikiko Nagashima and Peter F. Hitchcock, date: 12/20/2020, view: 1499, Q&A: 0
    [Abstract]

    Immunohistochemistry is a widely used technique to examine the expression and subcellular localization of proteins. This technique relies on the specificity of antibodies and requires adequate penetration of antibodies into tissues. The latter is especially challenging for thick specimens, such as embryos and other whole-mount preparations. Here

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    Multitarget Immunohistochemistry for Confocal and Super-resolution Imaging of Plant Cell Wall Polysaccharides
    Authors:  Kalina T. Haas, Methieu Rivière, Raymond Wightman and Alexis Peaucelle, date: 10/05/2020, view: 2509, Q&A: 0
    [Abstract] The plant cell wall (PCW) is a pecto-cellulosic extracellular matrix that envelopes the plant cell. By integrating extra-and intra-cellular cues, PCW mediates a plethora of essential physiological functions. Notably, it permits controlled and oriented tissue growth by tuning its local mechano-chemical properties. To refine our knowledge of these ...
    Hippocampal Unicellular Recordings and Hippocampal-dependent Innate Behaviors in an Adolescent Mouse Model of Alzheimer’s disease
    [Abstract] Transgenic mice have been used to make valuable contributions to the field of neuroscience and model neurological diseases. The simultaneous functional analysis of hippocampal cell activity combined with hippocampal dependent innate task evaluations provides a reliable experimental approach to detect fine changes during early phases of ...
    Semi-quantitative Determination of Protein Expression Using Immunohistochemistry Staining and Analysis: An Integrated Protocol
    Authors:  Alexandra R Crowe and Wei Yue, date: 12/20/2019, view: 5571, Q&A: 0
    [Abstract] Semi-quantitative IHC is a powerful method for investigating protein expression and localization within tissues. The semi-quantitative immunohistochemistry (IHC) involves using software such as free software ImageJ Fiji to conduct deconvolution and downstream analysis. Currently, there is lack of an integrated protocol that includes a detailed ...
    Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
    Authors:  Mengdie Wang, Gregory C. Rogers and Anne E. Cress, date: 10/20/2019, view: 2701, Q&A: 0
    [Abstract] Centrosome numerical abnormalities have been reported in a variety of tumors. Centrosome numbers in cancer cells display both inter-tumor and intra-tumor heterogeneity. The over production of centrosomes (centrosome amplification) is unique in cancer cells and is a promising target for therapy. Thus, a method to quantify centrosome numbers on a ...
    In situ Hybridization of Plant-parasitic Nematode Globodera pallida Juveniles to Detect Gene Expression
    Authors:  Joanna Kud, Nejra Solo, Allan Caplan, Joseph C. Kuhl, Louise-Marie Dandurand and Fangming Xiao, date: 09/20/2019, view: 2549, Q&A: 0
    [Abstract] In this study, we describe a standard whole mount in situ hybridization method which is used to determine the spatial-temporal expression pattern of genes from Globodera spp. Unlike more invasive radioactive labeling approaches, this technique is based on a safe, highly specific enzyme-linked immunoassay where a Digoxigenin ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 3440, Q&A: 0
    [Abstract] In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
    Quantification of Cutaneous Ionocytes in Small Aquatic Organisms
    Authors:  Garfield T. Kwan, Shane H. Finnerty, Nicholas C. Wegner and Martin Tresguerres, date: 05/05/2019, view: 3281, Q&A: 0
    [Abstract] Aquatic organisms have specialized cells called ionocytes that regulate the ionic composition, osmolarity, and acid/base status of internal fluids. In small aquatic organisms such as fishes in their early life stages, ionocytes are typically found on the cutaneous surface and their abundance can change to help cope with various metabolic and ...



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