Visualize Microtubule Dynamics in Live Cells
To visualize microtubule dynamics in live cells, Thermo Fisher Scientific has developed Invitrogen Tubulin Tracker Deep Red, a fluorescent molecule that selectively stains microtubules in live cells.
Regulation of microtubule dynamics, mechanics and function …
2021年8月18日 · Microtubules are essential, multitasking protein polymers that serve as structural elements in most eukaryotic cells. They are dynamic, and their dynamics are controlled to help organize the...
Microtubules and Microscopes: How the Development of Light …
The ER images were digitally processed to enhance their tubular structure and remove background staining of the plasma membrane. Microtubule images were processed to remove background of nonpolymerized labeled tubulin in the cell.
A live-cell marker to visualize the dynamics of stable microtubules ...
2023年3月2日 · Here, we present StableMARK (Stable Microtubule-Associated Rigor-Kinesin), a live-cell marker to visualize stable MTs with high spatiotemporal resolution. We demonstrate that a rigor mutant of Kinesin-1 selectively binds to stable MTs without affecting MT organization and organelle transport.
Fluorescence Methods for the Analysis of Microtubule…
2020年1月20日 · Using double immunofluorescent staining of endothelial cells, mutual arrangement of microtubules and actin stress fibers has been described as well as the positional relationship of microtubules and specific endothelial VE-cadherin cell-cell contacts .
Single-shot 20-fold expansion microscopy - Nature Methods
2024年10月11日 · Here, we present an ExM protocol that achieves ~20× expansion (yielding <20-nm resolution on a conventional microscope) in a single expansion step, achieving the performance of iterative...
Elongator is a microtubule polymerase selective for …
2 天之前 · The microtubule-containing pellet was resuspended in cold BRB80 (depolymerisation 1; 1/10 of the initial SN1-volume) and incubated on ice with occasional pipetting up-and-down over 20 min to mechanically assist microtubule depolymerization.
Blinking Fluorescent Probes for Tubulin Nanoscopy in Living and …
2021年11月4日 · In living cells, microtubule staining by probes 5 and 6 was weak, largely overshadowed by bright staining of cytoplasmic vesicles (Figure S1a). In contrast, all CTX-based probes stained mainly microtubules, and probe 3 (HMSiR-tubulin) was the brightest.
Structural insights into the interplay between microtubule
2025年1月5日 · The γ-tubulin ring complex (γ-TuRC) is a structural template for controlled nucleation of microtubules from α/β-tubulin heterodimers. At the cytoplasmic side of the yeast spindle pole body ...
Molecular characterization of the TUBG1 meshwork’s influence on ...
2 天之前 · To investigate the effect of impaired TUBG expression on microtubules, we expressed Cas9-GFP-TUBG1-sg in U2OS cells for seven days and used immunofluorescence staining with antibodies against α-tubulin (a microtubule marker) and TUBG (Fig. 2 A&B).
Visualizing the Actin and Microtubule Cytoskeletons at the B-cell ...
Microtubule plus-end binding proteins, as well as cortical plus-end capture proteins mediate physical interactions between the actin and microtubule cytoskeletons, which allow them to be reorganized in a coordinated manner.
Negative staining is a rapid, qualitative method for analyzing microtubule structure at the EM level. Because negative staining involves deposition of heavy atom stains, structural artifacts such as flattening of the cylindrical microtubule and opening up of …
Probes for Tubulin and Other Cytoskeletal Proteins—Section 11.2
The nuclear stain DAPI (D1306, D3571, D21490) binds tightly to purified tubulin in vitro without interfering with microtubule assembly or GTP hydrolysis. DAPI binds to tubulin at sites different from those of paclitaxel, colchicine and vinblastine, and its binding is accompanied by shifts in the absorption spectra and fluorescence enhancement.
Optimal conditions for fluorescence of proteins of the actin and tubulin cytoskeleton are based on: preserving cell structure; properties of individual cytoskeletal proteins and any antibodies to be used; background fluorescence.
Morphological Evidence for Novel Roles of Microtubules in …
In this study, to determine the function of microtubules in the fast activation of phagocytosis, we carefully observed the morphology of microtubules and found a dynamically rearranging microtubule network, even in the resting stage of the macrophages.
A novel immunofluorescence method to visualize microtubules in …
2017年11月15日 · Here, we developed a novel method to visualize selectively antiparallel microtubule overlaps in the midzone. When cells are air-dried before fixation, aligned α-tubulin staining is observed and colocalized with PRC1 in the center of the midzone of anaphase and telophase cells, suggesting that antiparallel microtubule overlaps can be visualized ...
Blinking Fluorescent Probes for Tubulin Nanoscopy in Living and …
2021年11月19日 · In living cells, microtubule staining by probes 5 and 6 was weak, largely overshadowed by bright staining of cytoplasmic vesicles (Figure S1a). In contrast, all CTX-based probes stained mainly microtubules, and probe 3 (HMSiR-tubulin) was the brightest.
Fluorescence Microscopy of Microtubules in Cultured Cells
Two experimental approaches based on fluorescence microscopy are commonly used to examine organization of microtubules in mammalian tissue culture cells. The first experimental approach involves indirect immunofluorescence staining of chemically fixed cells with tubulin antibody.
ViaFluor® Live Cell Microtubule Stains - Biotium
ViaFluor® Live Cell Microtubule Stains are cell-permeant probes for imaging the microtubule cytoskeleton in live cells. They are simple, rapid and sensitive stains that can be imaged without a wash step.
Microtubule acetylation is not required for HIV-1 infection or …
4 天之前 · Although microtubule acetylation is a well-studied post-translational modification in the context of cellular processes, its role during viral infections remains underexplored. ... Cells were transduced with the indicated aTAT1 construct and subjected to immunofluorescence staining for acetylated tubulin, alpha tubulin, and nuclear staining ...
Chapter 16 Studying Microtubules by Electron Microscopy
2008年1月1日 · Images of paclitaxel-stabilized microtubules visualized using negative stain EM. The microtubule lattice can be easily visualized by staining with uranyl acetate; protofilaments are indicated with dotted lines on the left image.
The where, when and how of microtubule nucleation - The …
2012年10月1日 · Microtubule polymerization occurs spontaneously in vitro, but under physiological conditions this process requires a nucleator that mimics or stabilizes a small microtubule seed formed from multiple α-tubulin–β-tubulin heterodimers.
Visualization of microtubules of cells in situ by indirect ...
Microtubule staining patterns can be visualized within cells in situ on the surface of fish scales from the squirrel fish, Holocentrus ascensionis, and the common goldfish, Carassius auratus, after incubation with antibodies to sea urchin tubulin and fluorescein-labeled goat antibodies to rabbit immunoglobulin G. Chromatophores in situ from both...
Staining Actin and Tubulin – from WB to Live Cell Imaging
2015年5月6日 · Spirochrome recently released unique fluorescent stains for monitoring both major components of thecytoskeleton easily in living cells. These fluorescent stains (SiR-Actin and SiR-Tubulin) are cell permeable compounds which stain F-actin (and microtubules, respectively; see Fig. 6 and Fig.7).