Protocols in Current Issue
    Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice
    Authors:  Hiroki Ueharu, Jingwen Yang, Yoshihiro Komatsu and Yuji Mishina, date: 04/05/2022, view: 454, Q&A: 0

    Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute

    Efficient Method to Differentiate Mouse Embryonic Stem Cells into Macrophages in vitro
    Authors:  Qimin Hai, Juying Han, Sophia Wells and Jonathan D. Smith, date: 02/05/2022, view: 1345, Q&A: 0

    Macrophages are key cells in the innate immune system and play a role in a variety of diseases. However, macrophages are terminally differentiated and difficult to manipulate genetically via transfection or through CRISPR-Cas9 gene editing. To overcome this limitation, we provide a simplified protocol for the generation of mouse embryonic stem

    Wholemount in situ Hybridization for Spatial-temporal Visualization of Gene Expression in Early Post-implantation Mouse Embryos
    Authors:  Xianfa Yang, Yingying Chen, Lu Song, Ting Zhang and Naihe Jing, date: 11/20/2021, view: 977, Q&A: 0

    Regionalized distribution of genes plays crucial roles in the formation of the spatial pattern in tissues and embryos during development. In situ hybridization has been one of the most widely used methods to screen, identify, and validate the spatial distribution of genes in tissues and embryos, due to its relative simplicity and low cost.

    Whole-mount Immunohistochemistry to Visualize Mouse Embryonic Dermal Lymphatic Vasculature
    Authors:  Xin Geng and R. Sathish Srinivasan, date: 10/20/2021, view: 957, Q&A: 0

    Lymphatic vessels are abundant in the skin where they regulate interstitial fluid uptake and immune surveillance. Defects in dermal lymphatic vessels, such as fewer vessels and abnormal lymphatic vessel coverage with mural cells, are frequently associated with lymphedema and other lymphatic disorders. Whole-mount immunohistochemistry allows the

    Treadmill Running of Mouse as a Model for Studying Influence of Maternal Exercise on Offspring
    Authors:  Song Ah Chae, Jun Seok Son, Mei-Jun Zhu, Jeanene M. de Avila and Min Du, date: 12/05/2020, view: 2262, Q&A: 0

    Epidemiological studies robustly show the beneficial effects of maternal exercise in reducing maternal birth complications and improving neonatal outcomes, though underlying mechanisms remain poorly understood. To facilitate mechanistic exploration, a protocol for maternal exercise of mice is established, with the regimen following the exercise

    Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules
    Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 2657, Q&A: 0
    [Abstract] Cells inside the body are subjected to various mechanical stress, such as stretch or compression provided by surrounding cells, shear stresses by blood or lymph flows, and normal stresses by luminal liquids. Force loading to the biological tissues is a fundamental method to better understand cellular responses to such mechanical stimuli. There ...
    Primed Track: Reliable Volumetric Single-cell Tracking and Lineage Tracing of Living Specimen with Dual-labeling Approaches
    Authors:  Maaike Welling, Konstantinos Kalyviotis and Periklis Pantazis, date: 06/05/2020, view: 2385, Q&A: 0
    [Abstract] Mammalian embryonic development starts with a single fertilized zygote that develops into a blastocyst embryo consisting of three cell types that evolve into either embryonic or extra-embryonic tissues. Lineage tracing of these cells can provide important information about the molecular and cellular dynamics contributing to fate allocation during ...
    RNA Interactome Identification via RNA-BioID in Mouse Embryonic Fibroblasts
    Authors:  Joyita Mukherjee, Mirita Franz-Wachtel, Boris Maček and Ralf-Peter Jansen, date: 01/05/2020, view: 3677, Q&A: 0
    [Abstract] Cytoplasmic localization of mRNAs is common to all organisms and serves the spatial expression of genes. Cis-acting RNA signals (mostly found in the mRNA’s 3'-UTR), called zipcodes recruit trans-acting RNA-binding proteins that facilitate the localization of the mRNA. UV-cross-linking or affinity purification has been applied to identify such ...
    Opto-magnetic Selection and Isolation of Single Cells
    Authors:  Loïc Binan, Joannie Roy and Santiago Costantino, date: 11/20/2019, view: 2903, Q&A: 0
    [Abstract] Capturing single cells from large heterogenous populations based solely on observable traits is necessary for many cell biology applications and remains a major technical challenge. The protocol we present allows the isolation of viable and metabolically active cells selected for their shape, migration speed, contact to other cells, or ...
    Explant Culture of the Embryonic Mouse Spinal Cord and Gene Transfer by ex vivo Electroporation
    [Abstract] Developing axons change responsiveness to guidance cues during the journey to synapse with target cells. Axon crossing at the ventral midline serves as a model for studying how axons accomplish such a switch in their response. Although primary neuron culture has been a versatile technique for elucidating various developmental mechanisms, many in ...

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