Membrane trafficking is central to cell plate construction during plant cytokinesis. Studies on cell plate formation can provide answers to basic biology questions including molecular mechanisms of membrane trafficking, tissue patterning, and cytoskeletal dynamics. Consequently, a detailed understanding of cytokinesis depends on the characterization of molecules that function in the formation, transport, targeting, and fusion of membrane vesicles and delivery of proteins to the developing and maturing plate. This chapter describes a pipeline based on fluorescence recovery after photobleaching (FRAP) to measure and analyze turnover of peripheral or transmembrane proteins on the cell plate. The approach described here can also be applied in other biological contexts.
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http://dx.doi.org/10.1007/978-1-0716-1744-1_14 | DOI Listing |
J Vis Exp
December 2024
Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile;
Plasmids play a vital role in synthetic biology by enabling the introduction and expression of foreign genes in various organisms, thereby facilitating the construction of biological circuits and pathways within and between cell populations. For many applications, maintaining functional plasmids without antibiotic selection is critical. This study introduces an open-hardware-based microfluidic workflow for analyzing plasmid retention by culturing single cells in gel microdroplets and quantifying microcolonies using fluorescence microscopy.
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January 2025
Laboratoire Interdisciplinaire de Physique (LIPhy), Université Grenoble Alpes, CNRS, Grenoble, France.
Cell-generated forces play a critical role in driving and regulating complex biological processes, such as cell migration and division and cell and tissue morphogenesis in development and disease. Traction force microscopy (TFM) is an established technique developed in the field of mechanobiology used to quantify cellular forces exerted on soft substrates and internal mechanical tissue stresses. TFM measures cell-generated traction forces in 2D or 3D environments with varying mechanical and biochemical properties.
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January 2025
Glycometabolic Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, Riken, 2-1 Hirosawa, Wako Saitama, Japan.
Cytosolic peptide:-glycanase (PNGase/NGLY1 in mammals), an amidase classified under EC:3.5.1.
View Article and Find Full Text PDFBiophys Physicobiol
September 2024
Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
Single-molecule imaging provides information on diffusion dynamics, oligomerization, and protein-protein interactions in living cells. To simultaneously monitor different types of proteins at the single-molecule level, orthogonal fluorescent labeling methods with different photostable dyes are required. G-protein-coupled receptors (GPCRs), a major class of drug targets, are prototypical membrane receptors that have been studied using single-molecule imaging techniques.
View Article and Find Full Text PDFEndosc Ultrasound
December 2024
Center of Excellence for Stem Cell and Cell Therapy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Introduction: EUS-guided fine-needle organoid creation (EUS-FNO) from pancreatic cancer (PC) has been increasingly important for precision medicine. The cost for pancreatic organoid creation is substantial and close to 2000 USD/specimen in our institution, and the specimen has to be processed immediately after tissue acquisition so the more passes and specimens, the higher cost of organoid creation will incur. To date, no prospective comparison trial has answered how many needle passes of EUS-FNO needed for a successful organoid creation.
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