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    Protocols in Current Issue
    Flow Cytometric Quantification of Fatty Acid Uptake by Mycobacterium tuberculosis in Macrophages
    Authors:  Evgeniya V. Nazarova, Maria Podinovskaia, David G. Russell and Brian C. VanderVen, date: 02/20/2018, view: 60, Q&A: 0
    Mycobacterium tuberculosis (Mtb) has evolved to assimilate fatty acids from its host. However, until recently, there was no reliable way to quantify fatty acid uptake by the bacteria during host cell infection. Here we describe a new method ...
    Registration and Alignment Between in vivo Functional and Cytoarchitectonic Maps of Mouse Visual Cortex
    Authors:  Jun Zhuang, Quanxin Wang, Marc Takeno and Jack Waters, date: 02/20/2018, view: 44, Q&A: 0
    This protocol describes a method for registration of in vivo cortical retinotopic map with cytochrome c oxidase (CO) labeled architectonic maps of the same mouse brain through the alignment of vascular fiducials. By recording surface blood ...
    Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
    Authors:  Umesh Goyal, Chitra Jaiswal and Malancha Ta, date: 02/20/2018, view: 106, Q&A: 0
    Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal tissue which is easily attainable, and a promising ...
    Sebinger Culture: A System Optimized for Morphological Maturation and Imaging of Cultured Mouse Metanephric Primordia
    Authors:  Mona Elhendawi and Jamie A. Davies, date: 02/20/2018, view: 49, Q&A: 0
    Here, we present a detailed protocol on setting up embryonic renal organ cultures using a culture method that we have optimised for anatomical maturation and imaging. Our culture method places kidney rudiments on glass in a thin film of medium, ...
    Preparation of Amyloid Fibril Networks
    Authors:  Mirren Charnley , Jay Gilbert, Owen G. Jones and Nicholas P. Reynolds, date: 02/20/2018, view: 76, Q&A: 0
    Networks of amyloid nanofibrils fabricated from common globular proteins such as lysozyme and β-lactoglobulin have material properties that mimic the extracellular microenvironment of many cell types. Cells cultured on such amyloid fibril networks ...
    Characterization of Amyloid Fibril Networks by Atomic Force Microscopy
    Authors:  Mirren Charnley , Jay Gilbert, Owen G. Jones and Nicholas P. Reynolds, date: 02/20/2018, view: 78, Q&A: 0
    Dense networks of amyloid nanofibrils fabricated from common globular proteins adsorbed to solid supports can improve cell adhesion, spreading and differentiation compared to traditional flat, stiff 2D cell culture substrates like Tissue Culture ...
    Flow Cytometric Quantification of Fatty Acid Uptake by Mycobacterium tuberculosis in Macrophages
    Authors:  Evgeniya V. Nazarova, Maria Podinovskaia, David G. Russell and Brian C. VanderVen, date: 02/20/2018, view: 60, Q&A: 0
    [Abstract] Mycobacterium tuberculosis (Mtb) has evolved to assimilate fatty acids from its host. However, until recently, there was no reliable way to quantify fatty acid uptake by the bacteria during host cell infection. Here we describe a new method to quantify fatty acid uptake by intracellular bacilli. We infect macrophages with Mtb ...
    Registration and Alignment Between in vivo Functional and Cytoarchitectonic Maps of Mouse Visual Cortex
    Authors:  Jun Zhuang, Quanxin Wang, Marc Takeno and Jack Waters, date: 02/20/2018, view: 44, Q&A: 0
    [Abstract] This protocol describes a method for registration of in vivo cortical retinotopic map with cytochrome c oxidase (CO) labeled architectonic maps of the same mouse brain through the alignment of vascular fiducials. By recording surface blood vessel pattern and sequential alignment at each step, this method overcomes the challenge imposed by ...
    Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
    Authors:  Umesh Goyal, Chitra Jaiswal and Malancha Ta, date: 02/20/2018, view: 106, Q&A: 0
    [Abstract] Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal tissue which is easily attainable, and a promising source of stem cells with no associated ethical concerns. MSCs have been isolated from different ...
    Sebinger Culture: A System Optimized for Morphological Maturation and Imaging of Cultured Mouse Metanephric Primordia
    Authors:  Mona Elhendawi and Jamie A. Davies, date: 02/20/2018, view: 49, Q&A: 0
    [Abstract] Here, we present a detailed protocol on setting up embryonic renal organ cultures using a culture method that we have optimised for anatomical maturation and imaging. Our culture method places kidney rudiments on glass in a thin film of medium, which results in very flat cultures with all tubules in the same image plane. For reasons not yet ...
    Preparation of Amyloid Fibril Networks
    Authors:  Mirren Charnley , Jay Gilbert, Owen G. Jones and Nicholas P. Reynolds, date: 02/20/2018, view: 76, Q&A: 0
    [Abstract] Networks of amyloid nanofibrils fabricated from common globular proteins such as lysozyme and β-lactoglobulin have material properties that mimic the extracellular microenvironment of many cell types. Cells cultured on such amyloid fibril networks show improved attachment, spreading and in the case of mesenchymal stem cells improved ...
    Characterization of Amyloid Fibril Networks by Atomic Force Microscopy
    Authors:  Mirren Charnley , Jay Gilbert, Owen G. Jones and Nicholas P. Reynolds, date: 02/20/2018, view: 78, Q&A: 0
    [Abstract] Dense networks of amyloid nanofibrils fabricated from common globular proteins adsorbed to solid supports can improve cell adhesion, spreading and differentiation compared to traditional flat, stiff 2D cell culture substrates like Tissue Culture Polystyrene (TCPS). This is due to the fibrous, nanotopographic nature of the amyloid fibril networks ...
    Extraction of DNA from Murine Fecal Pellets for Downstream Phylogenetic Microbiota Analysis by Next-generation Sequencing
    Authors:  Elien Eeckhout and Andy Wullaert, date: 02/05/2018, view: 117, Q&A: 0
    [Abstract] Mouse models are widely used to evaluate the potential impact of the gut microbial composition on health and disease. Standardized protocols for sampling and storing murine feces, as well as for extracting DNA from these fecal pellets are needed to limit experimental variation between different studies. Both efficient lysis of the microbiota and ...
    Extraction and Analysis of Pan-metabolome Polar Metabolites by Ultra Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS)
    Authors:  Dania M. Malik, Seth Rhoades and Aalim Weljie, date: 02/05/2018, view: 120, Q&A: 0
    [Abstract] Modern triple quadrupole mass spectrometers provide the ability to detect and quantify a large number of metabolites using tandem mass spectrometry (MS/MS). Liquid chromatography (LC) is advantageous, as it does not require derivatization procedures and a large diversity in physiochemical characteristics of analytes can be accommodated through a ...
    Immunoprecipitation of Tri-methylated Capped RNA
    Authors:  Karen E. Hayes, Jamie A. Barr, Mingyi Xie, Joan A. Steitz and Ivan Martinez, date: 02/05/2018, view: 185, Q&A: 0
    [Abstract] Cellular quiescence (also known as G0 arrest) is characterized by reduced DNA replication, increased autophagy, and increased expression of cyclin-dependent kinase p27Kip1. Quiescence is essential for wound healing, organ regeneration, and preventing neoplasia. Previous findings indicate that microRNAs (miRNAs) play an ...
    Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
    Authors:  Malek Chouchane and Marcos Romualdo Costa, date: 02/05/2018, view: 199, Q&A: 0
    [Abstract] Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in ...