Cell Biology

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    Protocols in Current Issue
    Measuring Protein Synthesis during Cell Cycle by Azidohomoalanine (AHA) Labeling and Flow Cytometric Analysis
    Authors:  Koshi Imami and Tomoharu Yasuda, date: 04/20/2019, view: 350, Q&A: 0
    Protein synthesis is one of the most fundamental biological processes to maintain cellular proteostasis. Azidohomoalaine (AHA) is a non-radioactive and “clickable” amino acid analog of methionine which can be incorporated into newly synthesized ...
    Immunofluorescence Staining of WT-1/Podocalyxin on Mouse Kidney Sections
    Authors:  Carole Henique, Olivia Lenoir and Pierre-Louis Tharaux, date: 04/20/2019, view: 228, Q&A: 0
    In glomerular diseases, podocytes are one type of cells involved in dysfunction of glomerular filtration. In these conditions, podocyte proteins expression change. Therefore, immunofluorescent staining of podocytes can be performed to visualize ...
    Preparation and Manipulation of Olfactory Epithelium Explant Cultures for Measurement of the Mechanical Tension of Individual Axons Using the Biomembrane Force Probe
    Authors:  Coralie Fouquet and Alain Trembleau, date: 04/20/2019, view: 111, Q&A: 0
    In this paper, we describe a protocol allowing measurement of the mechanical tension of individual axons grown ex vivo from neural tissue explants. This protocol was developed with primary cultures of olfactory epithelium explants from ...
    Time-lapse Whole-field Fluorescence Imaging of Microglia Processes Motility in Acute Mouse Hippocampal Slices and Analysis
    Microglia are the resident immune cells of the central nervous system (CNS). In the last year, the improvements in the transgenic mouse technologies and imaging techniques have shed light on microglia functions under physiological conditions. ...
    Measuring Protein Synthesis during Cell Cycle by Azidohomoalanine (AHA) Labeling and Flow Cytometric Analysis
    Authors:  Koshi Imami and Tomoharu Yasuda, date: 04/20/2019, view: 350, Q&A: 0
    [Abstract] Protein synthesis is one of the most fundamental biological processes to maintain cellular proteostasis. Azidohomoalaine (AHA) is a non-radioactive and “clickable” amino acid analog of methionine which can be incorporated into newly synthesized proteins. Thus, AHA-labeled nascent proteins can be detected and quantified through fluorescent labeling ...
    Immunofluorescence Staining of WT-1/Podocalyxin on Mouse Kidney Sections
    Authors:  Carole Henique, Olivia Lenoir and Pierre-Louis Tharaux, date: 04/20/2019, view: 228, Q&A: 0
    [Abstract] In glomerular diseases, podocytes are one type of cells involved in dysfunction of glomerular filtration. In these conditions, podocyte proteins expression change. Therefore, immunofluorescent staining of podocytes can be performed to visualize podocyte localization of proteins of interest. In this protocol, we detail a method to stain podocytes ...
    Preparation and Manipulation of Olfactory Epithelium Explant Cultures for Measurement of the Mechanical Tension of Individual Axons Using the Biomembrane Force Probe
    Authors:  Coralie Fouquet and Alain Trembleau, date: 04/20/2019, view: 111, Q&A: 0
    [Abstract] In this paper, we describe a protocol allowing measurement of the mechanical tension of individual axons grown ex vivo from neural tissue explants. This protocol was developed with primary cultures of olfactory epithelium explants from embryonic (E13.5) mice. It includes a detailed description of explant dissection and culture, as well as ...
    Time-lapse Whole-field Fluorescence Imaging of Microglia Processes Motility in Acute Mouse Hippocampal Slices and Analysis
    [Abstract] Microglia are the resident immune cells of the central nervous system (CNS). In the last year, the improvements in the transgenic mouse technologies and imaging techniques have shed light on microglia functions under physiological conditions. Microglia continuously scan the brain parenchyma with their highly motile processes, maintaining tissue ...
    Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy
    Author:  Mateusz Majda, date: 04/05/2019, view: 432, Q&A: 0
    [Abstract] Plant cell walls consist of different polysaccharides and structural proteins, which form a rigid layer located outside of the plasma membrane. The wall is also a very dynamic cell composite, which is characterized by complex polysaccharide interactions and various modifications during cell development. The visualization of cell wall components in ...
    Quantitative Plasmodesmata Permeability Assay for Pavement Cells of Arabidopsis Leaves
    Authors:  Min Diao, Qiannan Wang and Shanjin Huang, date: 04/05/2019, view: 417, Q&A: 0
    [Abstract] Plasmodesmata (PD) are intercellular channels between walled plant cells that enable the transportation of materials between adjacent cells, which are important for plant growth and development. The permeability of PD must be tightly regulated. Assays to determine the permeability of PD are crucial for related studies on the regulation of PD ...
    Delivering "Chromatic Bacteria" Fluorescent Protein Tags to Proteobacteria Using Conjugation
    Authors:  Rudolf O Schlechter and Mitja NP Remus-Emsermann, date: 04/05/2019, view: 300, Q&A: 0
    [Abstract] Recently, we published a large and versatile set of plasmids, the chromatic bacteria toolbox, to deliver eight different fluorescent protein genes and four combinations of antibiotic resistance genes to Gram-negative bacteria. Fluorescent tags are important tools for single-cell microbiology, synthetic community studies, biofilm, and host-microbe ...
    Straight Channel Microfluidic Chips for the Study of Platelet Adhesion under Flow
    Authors:  Alexander Dupuy, Lining Arnold Ju and Freda H Passam, date: 03/20/2019, view: 714, Q&A: 0
    [Abstract] Microfluidic devices have become an integral method of cardiovascular research as they enable the study of shear force in biological processes, such as platelet function and thrombus formation. Furthermore, microfluidic chips offer the benefits of ex vivo testing of platelet adhesion using small amounts of blood or purified platelets. ...
    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: 916, 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 ...
    Primary Embryonic Rat Cortical Neuronal Culture and Chronic Rotenone Treatment in Microfluidic Culture Devices
    Authors:  Victor S. Van Laar, Beth Arnold and Sarah B Berman, date: 03/20/2019, view: 432, Q&A: 0
    [Abstract] In the study of neurodegenerative diseases, it is imperative to study the cellular and molecular changes associated with pathogenesis in the relevant cell type, central nervous system neurons. The unique compartmentalized morphology and bioenergetic needs of primary neurons present complications for their study in culture. Recent microculture ...



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