Mammalia

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    Protocols in Current Issue
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 140, Q&A: 0
    Hepatic stellate cells (HSCs), alternatively known as liver pericytes, can differentiate into myofibroblasts and secrete extra-cellular matrix components, thereby promoting wound healing and fibrosis. Studying HSCs can provide insights into the ...
    Adoptive Transfer of Basophils Enriched from Mouse Spleen
    Authors:  Adrian M. Piliponsky and Stephen J. Galli, date: 11/05/2019, view: 66, Q&A: 0
    CD49b is a member of the integrin family, expressed on basophils, natural killer (NK) cells and a subset CD4+ T cells in the spleen. This protocol describes the adoptive transfer of basophil-enriched CD49b+ cells obtained from ...
    Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy
    Advances in fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) offer unprecedented opportunities for studying diverse proteins and nanostructures involved in fundamental cell biology. It is now possible to ...
    Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
    Authors:  Julia K. Doh, Caroline A. Enns and Kimberly E. Beatty, date: 11/05/2019, view: 86, Q&A: 0
    Genetically-encoded tags are useful tools for multicolor and multi-scale cellular imaging. Versatile Interacting Peptide (VIP) tags, such as VIPER, are new genetically-encoded tags that can be used in various imaging applications. VIP tags consist ...
    Generation of CoilR Probe Peptides for VIPER-labeling of Cellular Proteins
    Authors:  Julia K. Doh, Savannah J. Tobin and Kimberly E. Beatty, date: 11/05/2019, view: 97, Q&A: 0
    Versatile Interacting Peptide (VIP) tags are a new class of genetically-encoded tag designed for imaging cellular proteins by fluorescence and electron microscopy. In 2018, we reported the VIPER tag (Doh et al., 2018), which contains two ...
    High Throughput Traction Force Microscopy for Multicellular Islands on Combinatorial Microarrays
    Authors:  Ian C. Berg and Gregory H. Underhill, date: 11/05/2019, view: 114, Q&A: 0
    The composition and mechanical properties of the cellular microenvironment along with the resulting distribution of cellular devolved forces can affect cellular function and behavior. Traction Force Microscopy (TFM) provides a method to measure the ...
    Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid
    Authors:  Jeroen Witteveldt and Sara Macias, date: 11/05/2019, view: 81, Q&A: 0
    Controlled differentiation of embryonic stem cells is an essential tool in stem cell research. In this protocol, we describe a simple differentiation protocol involving the induction of embryoid body formation in mouse embryonic stem cells (mESC) ...
    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: 62, Q&A: 0
    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 ...
    Model of Chemotherapy-associated Mucositis and Oral Opportunistic Infections
    Authors:  Takanori Sobue, Martinna Bertolini, Angela Thompson and Anna Dongari-Bagtzoglou, date: 11/05/2019, view: 95, Q&A: 0
    Oral mucositis is a common complication of cancer chemotherapy treatment. Because of the lack of relevant oral mucositis experimental models, it is not clear how chemotherapeutic agents injure the oral mucosa and if commensal microorganisms ...
    In vitro Screening of Antileishmanial Activity of Natural Product Compounds: Determination of IC50, CC50 and SI Values
    Authors:  Olga S. Koutsoni, Kalliopi Karampetsou and Eleni Dotsika, date: 11/05/2019, view: 87, Q&A: 0
    Neglected tropical diseases gain the scientific interest of numerous research programs in an attempt to achieve their effective control or elimination. In this attempt, more cutting-edge public health policies and research are needed for the ...
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 140, Q&A: 0
    [Abstract] Hepatic stellate cells (HSCs), alternatively known as liver pericytes, can differentiate into myofibroblasts and secrete extra-cellular matrix components, thereby promoting wound healing and fibrosis. Studying HSCs can provide insights into the pathological mechanisms governing these processes. HSC isolation methods typically comprise of enzymatic ...
    Adoptive Transfer of Basophils Enriched from Mouse Spleen
    Authors:  Adrian M. Piliponsky and Stephen J. Galli, date: 11/05/2019, view: 66, Q&A: 0
    [Abstract] CD49b is a member of the integrin family, expressed on basophils, natural killer (NK) cells and a subset CD4+ T cells in the spleen. This protocol describes the adoptive transfer of basophil-enriched CD49b+ cells obtained from mouse spleens by magnetic enrichment. This protocol can be used to assess the contribution of ...
    Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy
    [Abstract] Advances in fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) offer unprecedented opportunities for studying diverse proteins and nanostructures involved in fundamental cell biology. It is now possible to visualize and quantify the spatial organization of cellular proteins and other macromolecules by FM, ...
    Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
    Authors:  Julia K. Doh, Caroline A. Enns and Kimberly E. Beatty, date: 11/05/2019, view: 86, Q&A: 0
    [Abstract] Genetically-encoded tags are useful tools for multicolor and multi-scale cellular imaging. Versatile Interacting Peptide (VIP) tags, such as VIPER, are new genetically-encoded tags that can be used in various imaging applications. VIP tags consist of a coiled-coil heterodimer, with one peptide serving as the genetic tag and the other (“probe ...
    Generation of CoilR Probe Peptides for VIPER-labeling of Cellular Proteins
    Authors:  Julia K. Doh, Savannah J. Tobin and Kimberly E. Beatty, date: 11/05/2019, view: 97, Q&A: 0
    [Abstract] Versatile Interacting Peptide (VIP) tags are a new class of genetically-encoded tag designed for imaging cellular proteins by fluorescence and electron microscopy. In 2018, we reported the VIPER tag (Doh et al., 2018), which contains two elements: a genetically-encoded peptide tag (i.e., CoilE) and a probe peptide (i.e., ...
    High Throughput Traction Force Microscopy for Multicellular Islands on Combinatorial Microarrays
    Authors:  Ian C. Berg and Gregory H. Underhill, date: 11/05/2019, view: 114, Q&A: 0
    [Abstract] The composition and mechanical properties of the cellular microenvironment along with the resulting distribution of cellular devolved forces can affect cellular function and behavior. Traction Force Microscopy (TFM) provides a method to measure the forces applied to a surface by adherent cells. Numerous TFM systems have been described in ...
    Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid
    Authors:  Jeroen Witteveldt and Sara Macias, date: 11/05/2019, view: 81, Q&A: 0
    [Abstract] Controlled differentiation of embryonic stem cells is an essential tool in stem cell research. In this protocol, we describe a simple differentiation protocol involving the induction of embryoid body formation in mouse embryonic stem cells (mESC) using hanging droplets, followed by differentiation into a neuronal lineage.
    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: 62, 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 ...
    Model of Chemotherapy-associated Mucositis and Oral Opportunistic Infections
    Authors:  Takanori Sobue, Martinna Bertolini, Angela Thompson and Anna Dongari-Bagtzoglou, date: 11/05/2019, view: 95, Q&A: 0
    [Abstract] Oral mucositis is a common complication of cancer chemotherapy treatment. Because of the lack of relevant oral mucositis experimental models, it is not clear how chemotherapeutic agents injure the oral mucosa and if commensal microorganisms accelerate tissue damage. We developed an organotypic oral mucosa model that mimics cellular responses ...
    In vitro Screening of Antileishmanial Activity of Natural Product Compounds: Determination of IC50, CC50 and SI Values
    Authors:  Olga S. Koutsoni, Kalliopi Karampetsou and Eleni Dotsika, date: 11/05/2019, view: 87, Q&A: 0
    [Abstract] Neglected tropical diseases gain the scientific interest of numerous research programs in an attempt to achieve their effective control or elimination. In this attempt, more cutting-edge public health policies and research are needed for the discovery of new, safer and effective drugs originated from natural products. Here, we describe protocols ...



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