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    Electroporation of Labeled Antibodies to Visualize Endogenous Proteins and Posttranslational Modifications in Living Metazoan Cell Types
    Authors:  Sascha Conic, Dominique Desplancq, László Tora and Etienne Weiss, date: 11/05/2018, view: 127, Q&A: 0
    [Abstract] The spatiotemporal localization of different intracellular factors in real-time and their detection in live cells are important parameters to understand dynamic protein-based processes. Therefore, there is a demand to perform live-cell imaging and to measure endogenous protein dynamics in single cells. However, fluorescent labeling of endogenous ...
    Optical Clearing and Index Matching of Tissue Samples for High-resolution Fluorescence Imaging Using SeeDB2
    Authors:  Meng-Tsen Ke and Takeshi Imai, date: 10/20/2018, view: 416, Q&A: 0
    [Abstract] Tissue clearing techniques are useful for large-scale three-dimensional fluorescence imaging of thick tissues. However, high-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we present a water-based optical clearing and mounting media, SeeDB2, which is ...
    6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans
    Authors:  Sarah-Lena Offenburger and Anton Gartner, date: 09/20/2018, view: 308, Q&A: 0
    [Abstract] The nematode Caenorhabditis elegans is a powerful genetic model that can be used to investigate neuronal death. Research using C. elegans has been crucial to characterize cell death programmes that are conserved in mammals. Many neuronal signaling components, such as those mediating dopaminergic neurotransmission, are preserved ...
    HCV Reporter System (Viral Infection-Activated Split-Intein-Mediated Reporter System) for Testing Virus Cell-to-cell Transmission ex-vivo
    Authors:  Fanfan Zhao, Ting Zhao, Libin Deng, Dawei Lv, Xiaolong Zhang, Xiaoyu Pan, Jun Xu and Gang Long, date: 08/05/2018, view: 566, Q&A: 0
    [Abstract] Hepatitis C virus (HCV) spread involves two distinct entry pathways: cell-free transmission and cell-to-cell transmission. Cell-to-cell transmission is not only an efficient way for viruses to spread but also an effective method for escaping neutralizing antibodies. We adapted the viral infection-activated split-intein-mediated reporter system ...
    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: 1476, 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. ...
    Generation of Luciferase-expressing Tumor Cell Lines
    Authors:  Todd V. Brennan, Liwen Lin, Xiaopei Huang and Yiping Yang, date: 04/20/2018, view: 2673, Q&A: 1
    [Abstract] Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track tumor development and quantify tumor burden in vivo. For this purpose, the introduction of genes that confer tumors with bioluminescent ...
    Quantification of Bacterial Twitching Motility in Dense Colonies Using Transmitted Light Microscopy and Computational Image Analysis
    [Abstract] A method was developed to allow the quantification and mapping of relative bacterial twitching motility in dense samples, where tracking of individual bacteria was not feasible. In this approach, movies of bacterial films were acquired using differential interference contrast microscopy (DIC), and bacterial motility was then indirectly quantified ...
    Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia
    Authors:  Yusuke Tomina and Daniel A. Wagenaar, date: 03/05/2018, view: 1297, Q&A: 0
    [Abstract] In this protocol, we introduce an effective method for voltage-sensitive dye (VSD) loading and imaging of leech ganglia as used in Tomina and Wagenaar (2017). Dissection and dye loading procedures are the most critical steps toward successful whole-ganglion VSD imaging. The former entails the removal of the sheath that covers neurons in the ...
    Measurement of Lysosomal Size and Lysosomal Marker Intensities in Adult Caenorhabditis elegans
    Authors:  Julie M. Huynh, Hope Dang and Hanna Fares, date: 02/05/2018, view: 1324, Q&A: 0
    [Abstract] Assays have been developed to study trafficking in various tissues of Caenorhabditis elegans. Adult C. elegans intestinal cells are large and have extensive endocytic networks, thus making them a good system for deciphering the endocytic pathway using live imaging techniques. However, the presence of auto-fluorescent gut granules ...
    Measuring Mitochondrial ROS in Mammalian Cells with a Genetically Encoded Protein Sensor
    [Abstract] Reactive oxygen species (ROS) are not only known for their toxic effects on cells, but they also play an important role as second messengers. As such, they control a variety of cellular functions such as proliferation, metabolism, differentiation and apoptosis. Thus, ROS are involved in the regulation of multiple physiological and ...