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    Quantification of Starch in Guard Cells of Arabidopsis thaliana
    Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 178, Q&A: 0
    [Abstract] In this protocol, we describe how to quantify starch in guard cells of Arabidopsis thaliana using the fluorophore propidium iodide and confocal laser scanning microscopy. This simple method enables monitoring, with unprecedented resolution, the dynamics of starch in guard cells.
    FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans
    Authors:  Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 138, Q&A: 0
    [Abstract] FRAP (Fluorescence Recovery After Photobleaching) is probably the most direct method to investigate the dynamics of molecules in living cells. Here, we describe FRAP to quantify membrane fluidity in C. elegans. Using FRAP, we have shown that cold, glucose and exogenous saturated fatty acids can decrease the fluidity of cellular membranes ...
    Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation
    Authors:  Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 128, Q&A: 0
    [Abstract] Immunocytochemistry of cultured cells is a common and effective technique for determining compositions and localizations of proteins within cellular structures. However, traditional cultured cell fixation and staining protocols are not effective in preserving cultured cell cytonemes, long specialized filopodia that are dedicated to morphogen ...
    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: 308, 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 ...
    Increasing the Membrane Permeability of a Fern with DMSO
    Author:  Marcelo Garcés, date: 06/20/2018, view: 235, Q&A: 0
    [Abstract] Cell membrane prevents the entrance of extra molecules (e.g., transcription and translation inhibitors) into the cell. For studying the physiological effects of transcription and translation inhibitors on Hymenophyllum caudiculatum fronds, we incubate fronds with 0.1% DMSO to test if this increases cell membrane permeability ...
    Transient Expression Assay in NahG Arabidopsis Plants Using Agrobacterium tumefaciens
    Authors:  Pepe Cana-Quijada, Eduardo R. Bejarano, Rosa Lozano-Durán and Tábata Rosas-Díaz, date: 06/20/2018, view: 338, Q&A: 0
    [Abstract] Agrobacterium-mediated transient expression has greatly contributed to research in molecular plant biology but has low efficiency and inconsistency in Arabidopsis thaliana (Arabidopsis). Here, we describe a simple, efficient and fast protocol to make transient gene expression in NahG Arabidopsis plants using ...
    Intracellular and Mitochondrial Reactive Oxygen Species Measurement in Primary Cultured Neurons
    [Abstract] Reactive oxygen species (ROS) are chemically reactive oxygen containing molecules. ROS consist of radical oxygen species including superoxide anion (O2•−) and hydroxyl radical (•OH) and non-radical oxygen species such as hydrogen peroxide (H2O2), singlet oxygen (O2). ROS are generated by ...
    Slow and Fast Desiccation of Single-cell Thick Fronds of Filmy Ferns
    Author:  Marcelo Garcés, date: 06/05/2018, view: 235, Q&A: 0
    [Abstract] Filmy ferns can desiccate and recover after rehydration to resume photosynthesis. Slow and fast desiccation rates were compared in filmy fern fronds to determine whether structural or physiological differences may occur between desiccation rates. Slow desiccation is considered to be more similar to natural conditions experienced by plants that ...
    In vitro Explant Cultures to Interrogate Signaling Pathways that Regulate Mouse Lung Development
    Authors:  Changfu Yao, Gianni Carraro and Barry R. Stripp, date: 05/20/2018, view: 417, Q&A: 0
    [Abstract] Early mouse lung development, including specification of primordia, patterning of early endoderm and determination of regional progenitor cell fates, is tightly regulated. The ability to culture explanted embryonic lung tissue provides a tractable model to study cellular interactions and paracrine factors that regulate these processes. We provide ...
    An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells
    Authors:  Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 730, Q&A: 0
    [Abstract] In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...