Fluorescence microscopy is one of the most important approaches in the cell biologist's toolbox for studying the mitotic spindle. In fact, many of the key insights into our understanding of mitosis have been enabled by the visualization of mitotic processes using fluorescence microscopy. Here, we summarize some of the important considerations for imaging mitosis using fluorescence microscopy. Because light can damage live cells, we emphasize the importance of minimizing cellular damage while obtaining informative images.
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http://dx.doi.org/10.1007/978-1-4939-3542-0_1 | DOI Listing |
Methods Mol Biol
January 2025
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Heraklion, Crete, Greece.
Lineage tracing based on modern live imaging approaches enables to visualize, reconstruct, and analyze the developmental history, fate, and dynamic behaviors of cells in vivo in a direct, comprehensive, and quantitative manner. Light-sheet fluorescence microscopy (LSFM) has greatly boosted lineage tracing efforts, because fluorescently labeled specimens can be imaged in their entirety, over long periods of time, with high spatiotemporal resolution and minimal photodamage. In addition, an increasing arsenal of commercial and open-source software solutions for cell and nuclei segmentation and tracking can be employed to convert data from pixel-based to object-based representations, and to reconstruct the lineages of cells in their native context as they organize in tissues, organs, and whole organisms.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Institut de Génomique Fonctionnelle de Lyon (IGFL), UMR5242, Ecole Normale Supérieure de Lyon (ENSL), CNRS, Université de Lyon, Lyon, France.
Bimolecular Fluorescence Complementation (BiFC) is a powerful molecular imaging method used to visualize protein-protein interactions (PPIs) in living cells or organisms. BiFC is based on the reassociation of hemi-fragments of a monomeric fluorescent protein upon spatial proximity. It is compatible with conventional light microscopy, providing a resolution that is constrained by the diffraction of light to around 250 nm.
View Article and Find Full Text PDFJ Org Chem
January 2025
Department of Chemistry, University of California, Irvine, California 92697-2025, United States.
Activating agents enable the efficient preparation of organozinc complexes from zinc metal and organohalides, but their mechanisms had been obscured by the heterogeneous nature of these systems. Fluorescence microscopy, with the sensitivity to detect surface reaction intermediates, reveals distinct activating mechanisms of widely used activation strategies: trimethylsilyl chloride, LiCl, DMSO, and Rieke zinc powder. The resulting development of mechanistic models provides a better understanding of the oxidative-addition-solubilization sequence in organozinc reagent formation and contains lessons for methods development.
View Article and Find Full Text PDFMicrosc Res Tech
January 2025
Department of Computer Science, Cihan University, Sulaimaniyah, Kurdistan Region, Iraq.
Waveguide evanescent field fluorescence microscopy (WEFF) is an evanescent-based microscopy that utilizes a confined thin film of light, around 100 nm, to image the plasma membrane of cells attached to a waveguide. Low photobleaching and low background besides its high axial resolution allows time-lapse imaging to investigate changes in cell morphology in the presence or absence of chemical agents. Both large field of view (FOV) and uniform illumination are very important while imaging cell-substrate contacts with an evanescent field.
View Article and Find Full Text PDFFront Chem
December 2024
Department of Algology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Introduction: Compression of the nerve root by a lumbar disc herniation can cause radiating pain in the lower limbs, and the nerve root decompression treatment may leave some patients with motor dysfunction and reduced sensory function. Studies have shown that nerve growth factor (NGF) can promote nerve growth and repair, but high doses, long duration, and immune response have become bottlenecks of its clinical application.
Methods: To overcome this obstacle, we developed Prussian blue (PBs) nanoparticles with the bio-delivery function and antioxidant effects of nanoenzymes.
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