Eb3 Protein Imaging
Eb3 Protein Imaging - Related Videos
11:15
Imaging Protein-protein Interactions in vivo
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Cited by 5 •
2010
This protocol describes how to image protein-protein interactions using a FRET-based proximity assay.
04:26
FLIM-FRET Imaging for Characterization of Protein-Protein Interactions in Live Bacteria
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2025
The video describes the FLIM-FRET imaging technique to determine the protein-protein interaction in live bacteria expressing cytoplasmic proteins labeled with fluorescent proteins, a donor eGFP, and acceptor mCherry. The combined technique also allows the quantification of the interacting proteins.
08:46
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
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Cited by 34 •
2015
A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented.
08:59
4D Imaging of Protein Aggregation in Live Cells
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Cited by 4 •
2013
Cellular viability depends on timely and efficient management of protein misfolding. Here we describe a method for visualizing the different potential fates of a misfolded protein: refolding, degradation, or sequestration in inclusions. We demonstrate the use of a folding sensor, Ubc9ts, for monitoring proteostasis and aggregation quality control in live cells using 4D microscopy.
10:25
High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells
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Cited by 5 •
2016
Protocols for Human Umbilical Vein Endothelial Cell (HUVEC) culture, transient transfection of fluorescently-labeled markers of microtubule growth, live-cell imaging and automated analysis of interphase microtubule growth dynamics are detailed.
FAQs
Fluorescently labeled EB3 selectively associates with the plus ends of microtubules while those ends polymerize. This selective localization concentrates fluorescence at actively growing regions rather than along the entire microtubule, producing mobile comet-like signals. The resulting pattern allows researchers to follow where growth occurs and how it changes within living cells over time.
The position, speed, and direction of an EB3 comet provide complementary measurements of microtubule behavior. Position identifies the changing location of a growing end, speed reflects its movement during polymerization, and direction describes the trajectory through the cell. Together, these measurements help quantify microtubule growth, stability, and interactions with cellular structures.
Live-cell imaging preserves the time-dependent behavior that static observations cannot capture. EB3 comets can be followed as they move, allowing researchers to examine how microtubule organization changes rather than recording only one structural state. This temporal information is especially relevant when studying cytoskeletal responses in cells, biomaterials, or genetically modified systems.
A typical workflow uses fluorescently labeled EB3 in living cells, followed by microscopy that records the fluorescent signals over time. Researchers then track comet position, speed, and direction to quantify microtubule behavior. The measurements can be related to cytoskeletal organization, stability, and interactions with cellular structures, depending on the experimental question.
Bioengineers can use this approach when they need to determine how a cellular system organizes or changes its microtubule network. In particular, measurements can evaluate the effects of biomaterials or genetic modifications on cytoskeletal behavior. The resulting data help connect engineered conditions with changes in microtubule growth, stability, and organization.
EB3 measurements support bioengineering studies of cell migration, intracellular transport, tissue organization, and engineered cellular systems. Tracking comet behavior supplies quantitative information about the cytoskeletal activity underlying these processes. Researchers can therefore compare microtubule behavior across cellular or engineered conditions and assess how changes in organization may relate to system-level performance.