The ability of fluorescence microscopy to study labeled structures like cells has now been empowered to deliver greater spatial and temporal resolutions that were not possible before, thanks to a new method developed by University of Illinois researcher Gabriel Popescu and Ru Wang from his lab. Using this method, they were able to study the critical process of cell transport dynamics at multiple spatial and temporal scales and reveal, for the first time, properties of diffusive and directed motion transport in living cells.
Popescu leads the Quantitative Light Imaging Laboratory at Illinois' Beckman Institute, while Wang of the lab is first author on the paper reporting the method in Physical Review Letters. The new approach, called dispersion-relation fluorescence spectroscopy (DFS), labels molecules of interest with a fluorophore whose motion, the researchers write, "gives rise to spontaneous fluorescence intensity fluctuations that are analyzed to quantify the governing mass transport dynamics. These data are characterized by the effective dispersion relation."
That ability to study the directed and diffusive transport characteristics of cellular dispersion through a wide range of temporal and spatial scales is more comprehensive than using just fluorescence microscopy. It provides more information than existing methods, such as fluorescence correlation spectroscopy (FCS), which is widely used for studying molecular transport and diffusion coefficients at a fixed spatial scale.
This study used DFS to focus on the cell cytoskeleton subunit actin and found that "the fluorescently labeled actin cytoskeleton exhibits active transport motion along a direction parallel to the fibers and diffusive on the perpendicular direction." Those results, the researchers said, describe at what scale and when directed versus diffusive motion is taking place in the cell.
"So for the first time we think we're able to tell those apart and the spatial scales at which each is dominant," Popescu said.
"Some traditional methods are good at measuring local transport and some are good at measuring the larger scales," Wang said. "Our method gives a fuller view of what happens inside the cell, to the patterns of traffic. So we can look at both the local scale and at larger scales, and ask at which scale the motion transitions from random to directed motion."
Popescu said the multiplicity of scales the method offers over techniques like fluorescence correlated spectroscopy is key. Such knowledge would be valuable for researchers interested in the basic science of cellular dynamics, as well as those working in biomedical research, such as in analysis of a drug's effect on the body. This technique can be used with current fluorescence microscopy methods.
"I think that the beauty of this method is that you can use a commercial fluorescent microscope that is found everywhere to collect and analyze data in a very simple way," Wang said. "You don't need complicated expertise. Everyone can use it."
The method relies on taking time-resolved sequential data from fluorescent spectroscopic microscopy images and transforming them using the Fourier transform. This computational method enables easier understanding of the image data, providing a different representation of the image. Taking advantage of the respective frequency domains of patterns in the data, as this method does, is especially useful for trying to understand cellular dynamics like transport.
"So for the first time we saw this universal transport behavior in a living system: a clear combination of diffusive transport, like Brownian motion, and directed, deterministic transport," Popescu said. "As a general trend, we found that diffusion is dominant at short scales and directed transport at large distances."
Beckman Institute for Advanced Science and Technology: http://www.beckman.uiuc.edu/
This press release was posted to serve as a topic for discussion. Please comment below. We try our best to only post press releases that are associated with peer reviewed scientific literature. Critical discussions of the research are appreciated. If you need help finding a link to the original article, please contact us on twitter or via e-mail.
Researchers were surprised by what they found when they sandwiched a drop of water between two layers of an unusual two-dimensional material called graphene.
Scientists at Cern are suggesting they could soon detect miniature black holes, proving the existence of parallel universes and disproving the big bang theory of the creation of the universe.
The Curiosity rover makes a detection of nitrogen compounds which provide further evidence that ancient Mars would have been a habitable world.
Wild animals can predict earthquakes several weeks before they strike, and motion-activated cameras that track their movements could be adopted in quake-prone countries as an affordable early warning system, scientists said on Tuesday.
GENEVA (Reuters) - Scientists at Europe's CERN research center have had to postpone the imminent relaunch of their refitted 'Big Bang' machine, the Large Hadron Collider, because of a short-circuit in the wiring of one of the vital magnets.
Images taken by NASA's Dawn spacecraft show that a mysterious bright spot on dwarf planet Ceres could be a plume of water spurting from a deep, icy crater
Using seismic vibrations from earthquakes around the world, they are figuring out what Earth looks like below the surface
In honor of a very special Pi Day, enjoy this map that explores the human-made and natural structures that come closest to a perfect circle
The moon has a more complex history than previously thought with at least nine subsurface layers, results from ground-penetrating radar aboard China’s Yutu lunar rover shows, scientists said on Thursday.
Scientists at the CERN physics research center said on Thursday the mystery dark matter that makes up 96 percent of the stuff of the universe will be a prime target for their souped-up Large Hadron Collider (LHC) in the coming years.