Developmental Biology

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    Protocols in Current Issue
    Detection of mRNA by Whole Mount in situ Hybridization and DNA Extraction for Genotyping of Zebrafish Embryos
    Authors:  Rachna Narayanan and Andrew C. Oates, date: 03/20/2019, view: 1148, Q&A: 0
    [Abstract] In situ hybridization is used to visualize the spatial distribution of gene transcripts in tissues and in embryos, providing important information about disease and development. Current methods involve the use of complementary riboprobes incorporating non-radioactive labels that can be detected by immunohistochemistry and coupled to ...
    A Functionally Robust Phenotypic Screen that Identifies Drug Resistance-associated Genes Using 3D Cell Culture
    Authors:  Sun-Young Lee and Mina J. Bissell, date: 11/20/2018, view: 1379, Q&A: 0
    [Abstract] Drug resistance is a major obstacle in cancer treatment: A case in point is the failure of cancer patients to respond to tyrosine kinase inhibitors (TKI) of EGFR, a receptor that is highly expressed in many cancers. Identification of the targets and delineation of mechanisms of drug resistance remain major challenges. Traditional pharmacological ...
    Electron Microscopic Detection of Proteins and Protein Complexes in Mammalian Cells Using APEX-tagged, Conditionally Stable Nanobodies
    Authors:  Thomas E. Hall, James Rae, Nicholas Ariotti and Robert G. Parton, date: 11/20/2018, view: 1063, Q&A: 0
    [Abstract] The recent development of 3D electron microscopic techniques for cells and tissues has necessitated the development of new methods for the detection of proteins and protein-complexes in situ. The development of new genetic tags, such as the ascorbate peroxidase, APEX, for electron microscopic detection of tagged proteins has expanded the ...
    Mosaic Labeling and 3-Dimensional Morphological Analysis of Single Cells in the Zebrafish Left-right Organizer
    Authors:  Agnik Dasgupta, Andrew E. Jacob and Jeffrey D. Amack, date: 11/20/2018, view: 1089, Q&A: 0
    [Abstract] A transient epithelial structure called the left-right organizer (LRO) establishes left-right asymmetry in vertebrate embryos. Developmental defects that alter LRO formation result in left-right patterning errors that often lead to congenital heart malformations. However, little is known about mechanisms that regulate individual cell behaviors ...
    Optical Clearing and Index Matching of Tissue Samples for High-resolution Fluorescence Imaging Using SeeDB2
    Authors:  Meng-Tsen Ke and Takeshi Imai, date: 10/20/2018, view: 2359, Q&A: 1
    [Abstract] Tissue clearing techniques are useful for large-scale three-dimensional fluorescence imaging of thick tissues. However, high-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we present a water-based optical clearing and mounting media, SeeDB2, which is ...
    Activation of Fibroblast Contractility via Cell-Cell Interactions and Soluble Signals
    Authors:  Neha Pincha, Dyuti Saha, Tanay Bhatt, Ravindra K. Zirmire and Colin Jamora, date: 09/20/2018, view: 1418, Q&A: 0
    [Abstract] The collagen contraction assay is an in vitro, three-dimensional method to determine the factor(s) affecting the contractile behavior of activated cells such as fibroblasts in either physiological or pathological scenarios. The collagen lattices/hydrogels are seeded with fibroblasts to mimic the interactions between these cells and their ...
    Live Confocal Imaging of Brachypodium Spikelet Meristems
    Author:  Devin Lee O'Connor, date: 09/20/2018, view: 1330, Q&A: 0
    [Abstract] Live confocal imaging of fluorescent reporters and stains in plant meristems provides valuable measurements of gene expression, protein dynamics, cell polarity, cell division, and growth. The spikelet meristem in the grass Brachypodium distachyon (Brachypodium) is well suited to live imaging because of the ease of dissection, small ...
    Muscle Function Assessment Using a Drosophila Larvae Crawling Assay
    Authors:  Yanina Post and Achim Paululat, date: 07/20/2018, view: 1472, Q&A: 0
    [Abstract] Here we describe a simple method to measure larval muscle contraction and locomotion behavior. The method enables the user to acquire data, without the necessity of using expensive equipment (Rotstein et al., 2018). To measure contraction and locomotion behaviour, single larvae are positioned at the center of a humidified Petri dish. ...
    Mouse Mammary Gland Whole Mount Preparation and Analysis
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 07/05/2018, view: 3647, Q&A: 0
    [Abstract] The mammary gland undergoes extensive remodeling during pregnancy and is also subject to neoplastic processes both of which result in histological changes of the gland epithelial structure. Since the mammary tree is a complex three-dimensional structure a method is needed that provides an overview of the entire gland. Whole mounts provide this ...
    Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation
    Authors:  Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 1484, Q&A: 0
    [Abstract] Immunocytochemistry of cultured cells is a common and effective technique for determining compositions and localizations of proteins within cellular structures. However, traditional cultured cell fixation and staining protocols are not effective in preserving cultured cell cytonemes, long specialized filopodia that are dedicated to morphogen ...



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