ArticleBiophysical journal2013
Measuring actin flow in 3D cell protrusions.
Article in Biophysical journal, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Mapping Protein Diffusion from the Plasma Membrane to the Nucleus: Insights from Fluorescence Correlation Spectroscopy.Biomolecules · 2026Review
- Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells.Journal of biological physics · 2026Article
- Spatial triple-correlation spectroscopy reveals heterotrimer dynamics in live cells.Biophysical journal · 2026Article
- Pair correlation microscopy of intracellular molecular transport.Nature protocols · 2025Review
- Using enhanced number and brightness to measure protein oligomerization dynamics in live cells.Nature protocols · 2019Article
- A cytoskeletal clutch mediates cellular force transmission in a soft, three-dimensional extracellular matrix.Molecular biology of the cell · 2017Article
- Electrically tunable lens speeds up 3D orbital tracking.Biomedical optics express · 2015Article
- Single cell visualization of transcription kinetics variance of highly mobile identical genes using 3D nanoimaging.Scientific reports · 2015Article
- Disentangling Membrane Dynamics and Cell Migration; Differential Influences of F-actin and Cell-Matrix Adhesions.PloS one · 2015Article
- Chemotactic responses of neural stem cells to SDF-1α correlate closely with their differentiation status.Journal of molecular neuroscience : MN · 2014Article
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Authors and funding
3 authors.
Funding
Abstract
Actin dynamics is important in determining cell shape, tension, and migration. Methods such as fluorescent speckle microscopy and spatial temporal image correlation spectroscopy have been used to capture high-resolution actin turnover dynamics within cells in two dimensions. However, these methods are not directly applicable in 3D due to lower resolution and poor contrast. Here, we propose to capture actin flow in 3D with high spatial-temporal resolution by combining nanoscale precise imaging by rapid beam oscillation and fluctuation spectroscopy techniques. To measure the actin flow along cell protrusions in cell expressing actin-eGFP cultured in a type I collagen matrix, the laser was orbited around the protrusion and its trajectory was modulated in a clover-shaped pattern perpendicularly to the protrusion. Orbits were also alternated at two positions closely spaced along the protrusion axis. The pair cross-correlation function was applied to the fluorescence fluctuation from these two positions to capture the flow of actin. Measurements done on nonmoving cellular protrusion tips showed no pair-correlation at two orbital positions indicating a lack of flow of F-actin bundles. However, in some protrusions, the pair-correlation approach revealed directional flow of F-actin bundles near the protrusion surface with flow rates in the range of ∼1 μm/min, comparable to results in two dimensions using fluorescent speckle microscopy. Furthermore, we found that the actin flow rate is related to the distance to the protrusion tip. We also observed collagen deformation by concomitantly detecting collagen fibers with reflectance detection during these actin motions. The implementation of the nanoscale precise imaging by rapid beam oscillation method with a cloverleaf-shaped trajectory in conjunction with the pair cross-correlation function method provides a quantitative way of capturing dynamic flows and organization of proteins during cell migration in 3D in conditions of poor contrast.
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