Cell Biology

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    Protocols in Current Issue
    Using Imaging Flow Cytometry to Characterize Extracellular Vesicles Isolated from Cell Culture Media, Plasma or Urine
    Authors:  John R. Woollard, Amrutesh Puranik, Kyra L. Jordan and Lilach O. Lerman, date: 11/05/2019, view: 59, Q&A: 0
    [Abstract] The ability to non-invasively detect specific damage to the kidney has been limited. Identification of extracellular vesicles released by cells, especially when under duress, might allow for monitoring and identification of specific cell types within the kidney that are stressed. We have adapted a previously published traditional flow cytometry ...
    Quantitation of TLR4 Internalization in Response to LPS in Thioglycollate Elicited Peritoneal Mouse Macrophages by Flow Cytometry
    Authors:  Katharina Richard, Stefanie N. Vogel and Darren J. Perkins, date: 09/20/2019, view: 305, Q&A: 0
    [Abstract] Internalization of cell surface Toll-like Receptor 4 (TLR4) is a convenient read-out to measure LPS dependent activation of the TRIF adaptor pathway. We here provide a protocol to quantify the LPS dependent internalization of TLR4 using thioglycollate-elicited peritoneal macrophages by flow cytometry.
    Detachment Procedure of Bacteria from Atmospheric Particles for Flow-cytometry Counting
    Authors:  Carolina M. Araya, Alberto Cazorla and Isabel Reche, date: 06/20/2019, view: 822, Q&A: 0
    [Abstract] The protocol separates bacteria from atmospheric particles, obtaining with greater precision their abundance in the atmospheric deposition. This procedure is similar to the one used to separate bacteria in streambed sediments. The detachment procedure consists of a chemical treatment with sodium pyrophosphate and Tween 20 and a physical treatment ...
    Variable Dose Analysis: A Novel High-throughput RNAi Screening Method for Drosophila Cells
    Authors:  Katarzyna Sierzputowska, Chris R. Baxter and Benjamin E. Housden, date: 12/20/2018, view: 1265, Q&A: 0
    [Abstract] Genetic screens are a powerful approach to identify previously uncharacterized genes involved in specific biological processes. Several technologies have been developed for high-throughput screens using reagents such as RNAi or CRISPR, and each approach is associated with specific advantages and disadvantages. Variable Dose Analysis (VDA), is an ...
    Ultrasound Guided Intra-thymic Injection to Track Recent Thymic Emigrants and Investigate T Cell Development
    [Abstract] To track recent thymic emigrants (RTEs) or study T cell development in the thymus, intra-thymic injection of a cellular tag or precursor cells for various T cell lineages is often desired. However, the traditional surgical approach to expose the thymus for intra-thymic injection is time-consuming and can cause a high level of pain and stress to ...
    In-vivo gp100-specific Cytotoxic CD8+ T Cell Killing Assay
    Authors:  Mahdia Benkhoucha, Nicolas Molnarfi, Elodie Belnoue, Madiha Derouazi and Patrice H Lalive, date: 11/20/2018, view: 2072, Q&A: 0
    [Abstract] Cytotoxic CD8+ T lymphocytes (CTLs) represent a crucial component of the adaptive immune system and play a prominent role in the anti-tumor immune responses of both mice and humans.

    Cytotoxic CD8+ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived ...
    Papain-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry
    Authors:  Hanadie Yousef, Cathrin J Czupalla, Davis Lee, Eugene C Butcher and Tony Wyss-Coray, date: 11/20/2018, view: 3225, Q&A: 0
    [Abstract] Brain endothelial cells (BECs) form the integral component of the blood-brain barrier (BBB) which separates the systemic milieu from the brain parenchyma and protects the brain from pathogens and circulating factors. In order to study BEC biology, it was of particular interest to establish a method that enables researchers to investigate and ...
    Osteoblast Sorting and Intracellular Staining of CXCL12
    Authors:  Weihuan Wang, Gurnoor Majhail, Cui Lui and Lan Zhou, date: 05/20/2018, view: 2782, Q&A: 0
    [Abstract] Osteoblasts are bone marrow endosteum-lining niche cells playing important roles in the regulation of hematopoietic stem cells by secreting factors and cell adhesion molecules. Characterization of primary osteoblasts has been achieved through culture of outgrowth of collagenase treated bone. Immunophenotyping and flow-based analysis of long bone ...
    Generation of Busulfan Chimeric Mice for the Analysis of T Cell Population Dynamics
    Authors:  Thea Hogan, Andrew Yates and Benedict Seddon, date: 12/20/2017, view: 4677, Q&A: 0
    [Abstract] This protocol was developed to generate chimeric mice in which T lymphocytes could be stratified by age on the basis of congenic marker expression. The conditioning drug busulfan is used to ablate host haematopoietic stem cells while leaving the peripheral immune system intact. Busulfan treatment is followed by bone marrow transplantation (BMT), ...
    Analysing Temporal Dynamics of T Cell Division in vivo Using Ki67 and BrdU Co-labelling by Flow Cytometry
    Authors:  Thea Hogan, Andrew Yates and Benedict Seddon, date: 12/20/2017, view: 5243, Q&A: 0
    [Abstract] This protocol was developed to increase the richness of information available from in vivo T cell proliferation studies. DNA labelling techniques such as BrdU incorporation allow precise control of label administration and withdrawal, so that the division history of a population can be tracked in detail over long timeframes (days-weeks). ...



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