Cell Biology

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    Protocols in Current Issue
    Staining the Germline in Live Caenorhabditis elegans: Overcoming Challenges by Applying a Fluorescent-dye Feeding Strategy
    Author:  K. Adam Bohnert, date: 11/05/2018, view: 171, Q&A: 0
    C. elegans provides a tractable model organism for studying germline cell biology. Microscopy experiments are relatively facile, as this worm is transparent and germline development can be observed in real-time using DIC microscopy and/or ...
    Generation of BMEC Lines and in vitro BMEC-HSPC Co-culture Assays
    Authors:  Peipei Guo and Shahin Rafii, date: 11/05/2018, view: 121, Q&A: 0
    Endothelial cells (ECs) sustain the self-renewal and regeneration of adult hematopoietic stem and progenitor cells (HSPCs) via deployment of EC-derived paracrine factors, termed as angiocrine factors. Generation of durable ex vivo vascular ...
    Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells
    Authors:  Gal Haimovich and Jeffrey E. Gerst, date: 11/05/2018, view: 290, Q&A: 1
    Transcription and RNA decay play critical roles in the process of gene expression and the ability to accurately measure cellular mRNA levels is essential for understanding this regulation. Here, we describe a single-molecule fluorescent in situ ...
    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: 135, Q&A: 0
    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 ...
    Generation of Stable Expression Mammalian Cell Lines Using Lentivirus
    Authors:  Neha Tandon, Kaushik N. Thakkar, Edward L LaGory, Yu Liu and Amato J Giaccia, date: 11/05/2018, view: 95, Q&A: 0
    Lentiviruses are used very widely to generate stable expression mammalian cell lines. They are used for both gene down-regulation (by using shRNA) or for gene up-regulation (by using ORF of gene of interest). The technique of generating stable cell ...
    Ex vivo Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples
    To assess oncogenic potential, classical transformation assays are based on cell line models. However, cell line based models do not reflect the complexity of human tissues. We thus developed an inducible expression system for gene expression in ex ...
    Eye Drops for Delivery of Bioactive Compounds and BrdU to Stimulate Proliferation and Label Mitotically Active Cells in the Adult Rodent Retina
    Authors:  Cindy L. Linn, Sarah E. Webster and Mark K. Webster, date: 11/05/2018, view: 90, Q&A: 0
    Eye drop treatments are typically used to apply drugs to the anterior structures of the eye. Recently, however, studies have demonstrated that eye drops can reach the retina in the back of the eye if pharmacological agents are carried in appropriate ...
    Staining the Germline in Live Caenorhabditis elegans: Overcoming Challenges by Applying a Fluorescent-dye Feeding Strategy
    Author:  K. Adam Bohnert, date: 11/05/2018, view: 171, Q&A: 0
    [Abstract] C. elegans provides a tractable model organism for studying germline cell biology. Microscopy experiments are relatively facile, as this worm is transparent and germline development can be observed in real-time using DIC microscopy and/or fluorescent transgenes. Despite these many tools, robust staining techniques for imaging germ cells ...
    Generation of BMEC Lines and in vitro BMEC-HSPC Co-culture Assays
    Authors:  Peipei Guo and Shahin Rafii, date: 11/05/2018, view: 121, Q&A: 0
    [Abstract] Endothelial cells (ECs) sustain the self-renewal and regeneration of adult hematopoietic stem and progenitor cells (HSPCs) via deployment of EC-derived paracrine factors, termed as angiocrine factors. Generation of durable ex vivo vascular niche that maintains EC identity and preserves the angiocrine profile of organ of origin offers ...
    Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells
    Authors:  Gal Haimovich and Jeffrey E. Gerst, date: 11/05/2018, view: 290, Q&A: 1
    [Abstract] Transcription and RNA decay play critical roles in the process of gene expression and the ability to accurately measure cellular mRNA levels is essential for understanding this regulation. Here, we describe a single-molecule fluorescent in situ hybridization (smFISH) method (as performed in Haimovich et al., 2017) that detects ...
    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: 135, 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 ...
    Generation of Stable Expression Mammalian Cell Lines Using Lentivirus
    Authors:  Neha Tandon, Kaushik N. Thakkar, Edward L LaGory, Yu Liu and Amato J Giaccia, date: 11/05/2018, view: 95, Q&A: 0
    [Abstract] Lentiviruses are used very widely to generate stable expression mammalian cell lines. They are used for both gene down-regulation (by using shRNA) or for gene up-regulation (by using ORF of gene of interest). The technique of generating stable cell lines using 3rd generation lentivirus is very robust and it typically takes about 1-2 ...
    Ex vivo Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples
    [Abstract] To assess oncogenic potential, classical transformation assays are based on cell line models. However, cell line based models do not reflect the complexity of human tissues. We thus developed an inducible expression system for gene expression in ex vivo human tissues, which maintain native tissue architecture, such as epithelia and ...
    Eye Drops for Delivery of Bioactive Compounds and BrdU to Stimulate Proliferation and Label Mitotically Active Cells in the Adult Rodent Retina
    Authors:  Cindy L. Linn, Sarah E. Webster and Mark K. Webster, date: 11/05/2018, view: 90, Q&A: 0
    [Abstract] Eye drop treatments are typically used to apply drugs to the anterior structures of the eye. Recently, however, studies have demonstrated that eye drops can reach the retina in the back of the eye if pharmacological agents are carried in appropriate vehicles. Here, we introduce an eye drop procedure to deliver a drug (PNU-282987), in combination ...
    Murine Pharmacokinetic Studies
    [Abstract] Murine pharmacokinetics (PK) represents the absorption, distribution, metabolism, and elimination of drugs from the body, which helps to guide clinical studies, ultimately resulting in more effective drug treatment. The purpose of this protocol is to describe a serial bleeding protocol, obtaining blood samples at six time points from single mouse ...
    Vascular Permeability Assay in Human Coronary and Mouse Brachiocephalic Arteries
    [Abstract] Coronary artery disease remains an important cause of morbidity and mortality. Previous work, including ours, has focused on the role of intraplaque hemorrhage, particularly from immature microvessel angiogenesis, as an important contributor to plaque progression via increases in vascular permeability leading to further intraplaque hemorrhage, ...
    Examining Autophagy in Plant by Transmission Electron Microscopy (TEM)
    Authors:  Xiyin Zheng, Chenguang Zhao and Yule Liu, date: 10/20/2018, view: 297, Q&A: 0
    [Abstract] In plants, macroautophagy, here referred as autophagy, is a degradation pathway during which the double-membrane structure named autophagosome engulfs the cargo and then fuses with vacuole for material recycling.

    To investigate the process of autophagy, transmission electron microscopy (TEM) was used to monitor the ultrastructure of ...



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