Cell Biology

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    Protocols in Current Issue
    Calcium Imaging of Neuronal Activity under Gradually Changing Odor Stimulation in Caenorhabditis elegans
    Authors:  Yuki Tanimoto and Koutarou D. Kimura, date: 01/05/2021, view: 333, Q&A: 0

    Olfactory behavior is among the most fundamental animal behaviors both in the wild and in the laboratory. To elucidate the neural mechanisms underlying olfactory behavior, it is critical to measure neural responses to odorant concentration changes

    ...
    Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
    Authors:  Kawssar Harb, Michele Bertacchi and Michèle Studer, date: 01/05/2021, view: 608, Q&A: 0

    The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded

    ...
    Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
    Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 388, Q&A: 0

    The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process

    ...
    Measuring Bone Volume at Multiple Densities by Micro-computed Tomography
    Authors:  Emma C. Walker, Narelle E. McGregor, Audrey S. M. Chan and Natalie A. Sims, date: 01/05/2021, view: 610, Q&A: 0

    Bone strength is controlled by both bone mass, and the organization and quality of the bone material. The current standard method for measuring bone mass in mouse and rat studies is micro-computed tomography. This method typically uses a single

    ...
    A Simple Microplate Assay for Reactive Oxygen Species Generation and Rapid Cellular Protein Normalization
    Authors:  Neville S. Ng and Lezanne Ooi, date: 01/05/2021, view: 707, Q&A: 0
    2',7'-dichlorofluorescein (DCF) and derivatives are commonly used as fluorescent indicators of a broad spectrum of reactive oxygen species (ROS) generation in cell-based assays. However, there are numerous challenges inherent to the utilization of ...
    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 503, Q&A: 0

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a

    ...
    Stable Transformation of Arabidopsis thaliana Cell Suspension Cultures: A Case Study for the Overexpression of The COI1 Receptor
    Cell suspension cultures have been studied for decades to produce natural molecules. However, the difficulty in generating stably transformed cell lines has limited their use to produce high value chemicals reproducibly and in elevated quantities. ...
    A Parkinson’s Disease-relevant Mitochondrial and Neuronal Morphology High-throughput Screening Assay in LUHMES Cells

    Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease

    ...
    A Rigorous Quantitative Approach to Analyzing Phagocytosis Assays
    Authors:  Michael D. Caponegro, Kaitlyn Koenig Thompson, Maryam Tayyab and Stella E. Tsirka, date: 01/05/2020, view: 1334, Q&A: 0
    Studying monocytic cells in isolated systems in vitro contributes significantly to the understanding of innate immune physiology. Functional assays produce read outs which can be used to measure responses to selected stimuli, such as ...
    A Parkinson’s Disease-relevant Mitochondrial and Neuronal Morphology High-throughput Screening Assay in LUHMES Cells
    [Abstract]

    Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease is mitochondrial dysfunction, and much work has aimed to identify therapeutic compounds which can

    ...
    Stable Transformation of Arabidopsis thaliana Cell Suspension Cultures: A Case Study for the Overexpression of The COI1 Receptor
    [Abstract] Cell suspension cultures have been studied for decades to produce natural molecules. However, the difficulty in generating stably transformed cell lines has limited their use to produce high value chemicals reproducibly and in elevated quantities.

    In this protocol, a method to stably transform and maintain Arabidopsis cell ...
    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 503, Q&A: 0
    [Abstract]

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a transient 293T cell transfection-based cell adhesion assay to study selective cell adhesion of a

    ...
    A Simple Microplate Assay for Reactive Oxygen Species Generation and Rapid Cellular Protein Normalization
    Authors:  Neville S. Ng and Lezanne Ooi, date: 01/05/2021, view: 707, Q&A: 0
    [Abstract] 2',7'-dichlorofluorescein (DCF) and derivatives are commonly used as fluorescent indicators of a broad spectrum of reactive oxygen species (ROS) generation in cell-based assays. However, there are numerous challenges inherent to the utilization of DCF probes for intracellular microscopic analysis, including bleaching and probe efflux. Plate ...
    Measuring Bone Volume at Multiple Densities by Micro-computed Tomography
    Authors:  Emma C. Walker, Narelle E. McGregor, Audrey S. M. Chan and Natalie A. Sims, date: 01/05/2021, view: 610, Q&A: 0
    [Abstract]

    Bone strength is controlled by both bone mass, and the organization and quality of the bone material. The current standard method for measuring bone mass in mouse and rat studies is micro-computed tomography. This method typically uses a single threshold to identify bone material in the cortical and trabecular regions. However, this single

    ...
    Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
    Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 388, Q&A: 0
    [Abstract]

    The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain

    ...
    Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
    Authors:  Kawssar Harb, Michele Bertacchi and Michèle Studer, date: 01/05/2021, view: 608, Q&A: 0
    [Abstract]

    The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection

    ...
    Calcium Imaging of Neuronal Activity under Gradually Changing Odor Stimulation in Caenorhabditis elegans
    Authors:  Yuki Tanimoto and Koutarou D. Kimura, date: 01/05/2021, view: 333, Q&A: 0
    [Abstract]

    Olfactory behavior is among the most fundamental animal behaviors both in the wild and in the laboratory. To elucidate the neural mechanisms underlying olfactory behavior, it is critical to measure neural responses to odorant concentration changes resembling those that animals actually sense during olfactory behavior. However, reproducing the

    ...
    Lipid Droplet Isolation from Arabidopsis thaliana Leaves
    Authors:  Yovanny Izquierdo, Rubén Fernández-Santos, Tomás Cascón and Carmen Castresana, date: 12/20/2020, view: 555, Q&A: 0
    [Abstract]

    Lipid droplets (LDs) are neutral lipid aggregates surrounded by a phospholipid monolayer and specific proteins. In plants, they play a key role as energy source after seed germination, but are also formed in vegetative tissues in response to developmental or environmental conditions, where their functions are poorly understood. To elucidate these,

    ...
    Pea Aphid Rearing, Bacterial Infection and Hemocyte Phagocytosis Assay
    Authors:  Li Ma, Lu Liu and Zhiqiang Lu, date: 12/20/2020, view: 400, Q&A: 0
    [Abstract]

    Insects rely on the simple but effective innate immune system to combat infection. Cellular and humoral responses are interconnected and synergistic in insects’ innate immune system. Phagocytosis is one major cellular response. It is difficult to collect clean hemolymph from the small insect like pea aphid. Here, we provide a practicable

    ...



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