Immunology

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    Protocols in Current Issue
    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 ...
    Characterization of Mouse Adult Testicular Macrophage Populations by Immunofluorescence Imaging and Flow Cytometry
    Authors:  Noushin Mossadegh-Keller and Michael H. Sieweke, date: 03/05/2019, view: 672, Q&A: 0
    [Abstract] Testicular macrophages (tMΦ) are the most abundant immune cells residing in the testis, an immune-privileged organ. TMΦ are known to exhibit different functions, such as protecting spermatozoa from auto-immune attack by producing immunosuppressive cytokines and trophic roles in supporting spermatogenesis and male sex hormone production. They also ...
    Immunohistochemical Staining of TLR4 in Human Skeletal Muscle Samples
    Authors:  Daniela Verzola, Samantha Milanesi, Francesca Costigliolo and Giacomo Garibotto, date: 01/20/2019, view: 817, Q&A: 0
    [Abstract] Growing evidence suggests the involvement of TLR4, a receptor in the innate immune system, in muscle loss in uremia. Recently, we have evaluated TLR4 in human skeletal muscle from chronic kidney disease patients, by immunohistochemistry and image analysis. Unlike the commonly-used Western blot method, immunohistochemistry allows for the ...
    Quantification of Extracellular Double-stranded RNA Uptake and Subcellular Localization Using Flow Cytometry and Confocal Microscopy
    Authors:  Tan A Nguyen, Lachlan Whitehead and Ken C Pang, date: 06/20/2018, view: 1879, Q&A: 0
    [Abstract] Double-stranded RNA is a potent pathogen-associated molecular pattern (PAMP) produced as a by-product of viral replication and a well-known hallmark of viral infection. Viral dsRNAs can be released from infected cells into the extracellular space and internalized by neighboring cells via endocytosis. Mammals possess multiple pattern recognition ...
    Immunohistochemical Identification of Human Skeletal Muscle Macrophages
    [Abstract] Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage ...
    Measuring the Endocytic Recycling of Amyloid Precursor Protein (APP) in Neuro2a Cells
    Authors:  Florent Ubelmann, Tatiana Burrinha and Claudia Guimas Almeida, date: 12/05/2017, view: 2962, Q&A: 0
    [Abstract] The established primary trigger of Alzheimer’s disease’s is β-amyloid (Aβ) (Mucke and Selkoe, 2012). Amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes also depends on its recycling back to the plasma membrane (Koo et al., 1996; Ubelmann et ...
    Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction
    Authors:  Roberto Mendez and Santanu Banerjee, date: 11/05/2017, view: 3975, Q&A: 0
    [Abstract] In this protocol, we describe proximal ligation assay (PLA), an antibody-based detection method for protein-protein interaction. This method relies on specific binding of individual primary antibodies to the two putative interacting proteins. The primary antibodies need to have different hosts. The secondary antibodies against the two hosts have ...
    Macrophage Survival Assay Using High Content Microscopy
    Authors:  Remzi Onur Eren and Nicolas Fasel, date: 08/20/2017, view: 3247, Q&A: 0
    [Abstract] Macrophages maintain tissue homoeostasis by regulating inflammation and tissue repair mechanisms. Thus, the fate of macrophages has an impact on the state of the tissue. The aim of this protocol is to quantify macrophage survival using high content microscopy and image processing software. Here, we describe a high-content image based protocol to ...
    Functional ex-vivo Imaging of Arterial Cellular Recruitment and Lipid Extravasation
    [Abstract] The main purpose of this sophisticated and highly versatile method is to visualize and quantify structural vessel wall properties, cellular recruitment, and lipid/dextran extravasation under physiological conditions in living arteries. This will be of interest for a broad range of researchers within the field of inflammation, hypertension, ...
    In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
    Authors:  Alex Hopke and Robert T. Wheeler, date: 04/05/2017, view: 4203, Q&A: 0
    [Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell ...



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