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    Freeze-fracture-etching Electron Microscopy for Facile Analysis of Yeast Ultrastructure
    Authors:  Takuma Tsuji and Toyoshi Fujimoto, date: 09/20/2017, view: 98, Q&A: 0
    [Abstract] We describe a streamlined method that enables the quick observation of yeast ultrastructure by electron microscopy (EM). Yeast cells are high-pressure frozen, freeze-fractured to cut across the cytoplasm, and freeze-etched to sublimate ice in the cytosol and the organelle lumen. The cellular structures delineated by these procedures are coated by ...
    Uptake Assays to Monitor Anthracyclines Entry into Mammalian Cells
    Authors:  Nicolas Brosseau, Emil Andreev and Dindial Ramotar, date: 09/20/2017, view: 61, Q&A: 0
    [Abstract] Anthracyclines, such as doxorubicin and daunorubicin, are DNA damaging agents that autofluoresce and can be readily detected in cells. Herein, we developed suitable assays to quantify and localize daunorubicin in mammalian cells. These assays can be exploited to identify components that are involved in the uptake of anthracyclines.
    Peroxisome Motility Measurement and Quantification Assay
    Authors:  Jeremy Metz, Inês G. Castro and Michael Schrader, date: 09/05/2017, view: 215, Q&A: 0
    [Abstract] Organelle movement, distribution and interaction contribute to the organisation of the eukaryotic cell. Peroxisomes are multifunctional organelles which contribute to cellular lipid metabolism and ROS homeostasis. They distribute uniformly in mammalian cells and move along microtubules via kinesin and dynein motors. Their metabolic cooperation ...
    Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures
    Authors:  Jennifer Jung and Christian Behrends, date: 09/05/2017, view: 263, Q&A: 0
    [Abstract] Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy ...
    Determination of Local pH Differences within Living Salmonella Cells by High-resolution pH Imaging Based on pH-sensitive GFP Derivative, pHluorin(M153R)
    Authors:  Yusuke V. Morimoto, Nobunori Kami-ike, Keiichi Namba and Tohru Minamino, date: 09/05/2017, view: 169, Q&A: 0
    [Abstract] The bacterial flagellar type III protein export apparatus is composed of a transmembrane export gate complex and a cytoplasmic ATPase complex. The export apparatus requires ATP hydrolysis and the proton motive force across the cytoplasmic membrane to unfold and transport flagellar component proteins for the construction of the bacterial flagellum ...
    Multicolor Stimulated Emission Depletion (STED) Microscopy to Generate High-resolution Images of Respiratory Syncytial Virus Particles and Infected Cells
    [Abstract] Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these ...
    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 302, Q&A: 0
    [Abstract] We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...
    Staining of Membrane Receptors with Fluorescently-labeled DNA Aptamers for Super-resolution Imaging
    Authors:  Maria Angela Gomes de Castro, Claudia Höbartner and Felipe Opazo, date: 09/05/2017, view: 249, Q&A: 0
    [Abstract] One of the most prominent applications of fluorescent super-resolution microscopy is the study of nanodomain arrangements of receptors and the endocytic pathway. Staining methods are becoming crucial for answering questions on the nanoscale, therefore, the use of small and monovalent affinity probes is of great interest in super-resolution ...
    [Bio101] Leaf Clearing Protocol to Observe Stomata and Other Cells on Leaf Surface
    Author:  Nidhi Sharma, date: 09/05/2017, view: 157, Q&A: 0
    [Abstract] In this protocol, leaves are cleared and fixed in an ethanol and acetic acid solution, and mounted in Hoyer’s solution. The cleared leaves are imaged under differential interference contrast (DIC) microscope. This protocol is beneficial for studying stomata, hair cells, and other epidermal cells in plants.
    Macrophage Survival Assay Using High Content Microscopy
    Authors:  Remzi Onur Eren and Nicolas Fasel, date: 08/20/2017, view: 377, 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 ...