Fixed-cell imaging —Cell imaging —Cell Biology

Quantitative Analysis of Actin Cable Length in Yeast

Authors: Shane G. McInally, Jane Kondev and Bruce L. Goode, date: 05/05/2022, view: 781, Q&A: 0

[Abstract]

Polarized actin cables in S. cerevisiae are linear bundles of crosslinked actin filaments that are assembled by two formins, Bnr1 (localized to the bud neck), and Bni1 (localized to the bud tip). Actin is polymerized at these two sites, which results in cables extending along the cell cortex toward the back of the mother cell. These cables serve

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High Throughput Blood-brain Barrier Organoid Generation and Assessment of Receptor-Mediated Antibody Transcytosis

Authors: Elena Kassianidou, Claire Simonneau, Alina Gavrilov and Roberto Villaseñor, date: 04/20/2022, view: 551, Q&A: 0

[Abstract]

Targeting receptor-mediated transcytosis (RMT) is a successful strategy for drug delivery of biologic agents across the blood-brain barrier (BBB). The recent development of human BBB organoid models is a major advancement to help characterize the mechanisms of RMT and thus accelerate the design of brain delivery technologies. BBB organoids exhibit

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Gamete Fusion Assay in Mice

Author: Naokazu Inoue, date: 11/20/2021, view: 1127, Q&A: 0

[Abstract]

Gamete fusion, which is the final event of fertilization, is a crucial physiological event in the creation of a new fetus. In mammals, sperm IZUMO1 and oocyte IZUMO1R (JUNO) recognition play a role in triggering this process. Gamete fusion occurs through a complex but steady and unfailing intermolecular reaction because fertilization must ensure

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Phospho-protein Analysis in Adherent Cells Using Flow Cytometry

Authors: Renu Sharma, Amit Sharma, Atul Kumar and Bithiah Grace Jaganathan, date: 10/20/2019, view: 3921, Q&A: 0

[Abstract] Protein phosphorylation is one of the most important post-translational modifications, which acts as a reversible on or off switch for the activity of a large number of proteins. Analyzing the phosphorylation status of different proteins can reveal the alterations in the state of the cells in response to cellular damage, cancer and pharmaceutical ...

Cell-based Assay for Recruitment of DDR1 to Collagen-coated Beads

Authors: Victoria Juskaite and Birgit Leitinger, date: 08/20/2019, view: 3511, Q&A: 0

[Abstract] The discoidin domain receptors, DDR1 and DDR2, are key signaling receptors for the extracellular matrix protein collagen. The interactions of cells with collagen are difficult to study because of the difficulty to obtain native collagen fibers for in vitro studies. Thus, in vitro studies often use acid-soluble collagens in the ...

Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation

Authors: Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 4978, 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 ...

Tracking Endocytosis and Intracellular Trafficking of Epitope-tagged Syntaxin 3 by Antibody Feeding in Live, Polarized MDCK Cells

Authors: Adrian J. Giovannone, Elena Reales, Pallavi Bhattaram, Alberto Fraile-Ramos and Thomas Weimbs, date: 02/05/2018, view: 7286, Q&A: 0

[Abstract] The uptake and trafficking of cell surface receptors can be monitored by a technique called ‘antibody-feeding’ which uses an externally applied antibody to label the receptor on the surface of cultured, live cells. Here, we adapt the traditional antibody-feeding experiment to polarized epithelial cells (Madin-Darby Canine Kidney) grown on ...

The RiboPuromycylation Method (RPM): an Immunofluorescence Technique to Map Translation Sites at the Sub-cellular Level

Authors: Amandine Bastide, Jonathan W. Yewdell and Alexandre David, date: 01/05/2018, view: 10540, Q&A: 0

[Abstract] While isotopic labeling of amino acids remains the reference method in the field for quantifying translation rate, it does not provide any information on spatial localization of translation sites. The rationale behind developing the ribopuromycylation method (RPM) was primarily to map translation sites at the sub-cellular level while avoiding ...

Immunogold Localization of Molecular Constituents Associated with Basal Bodies, Flagella, and Extracellular Matrices in Male Gametes of Land Plants

Authors: Renee A. Lopez, Katayoun Mansouri, Jason S. Henry, Nicholas D. Flowers, Kevin C. Vaughn and Karen Sue Renzaglia, date: 11/05/2017, view: 5852, Q&A: 0

[Abstract] Male gametes (spermatozoids) are the only motile cells produced during the life cycle of land plants. While absent from flowering and most cone-bearing plants, motile cells are found in less derived taxa, including bryophytes (mosses, liverworts and hornworts), pteridophytes (lycophytes and ferns) and some seed plants (Ginkgo and cycads). ...

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: 7731, Q&A: 3

[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 ...

Cell Biology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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