"With more traditional visualization methods, you can't see the level of detail you need. It's important to look at not only what's present, but also when and where it's present in the cell," Timlin said.
The technique used by Timlin and Aaron builds on superresolution capabilities developed in recent years, but goes another step by adding dual-color capabilities to the relatively new stochastic optical reconstruction microscopy, or STORM. The combination enables the Sandia team to get a more complete picture by simultaneously imaging LPS and TLR4 receptors on the membrane.
"Current light microscopy capabilities are akin to looking out the window of an airplane and seeing the irrigation circles. You know that plants are there, but you can't tell what kinds of plants they are or what shape the leaves are," said Carson, a Sandia immunologist who was an integral part of the project. "But with this technology, it's like zooming in and seeing the leaves and the structure of the plants. That buys you a lot in terms of understanding what's happening within a cell and specifically how the proteins involved interact."
In 2009, the National Institutes of Health awarded Timlin a five-year, $300,000-a-year innovation grant. Next on the team's agenda is developing the capability to image live cells in real time using spectral Stimulated Emission Depletion, or STED. "We're working toward using a version of superresolution that's much more live-cell friendly, and extending that in terms of what colors are available to do multiple colors, while maintaining the live-cell friendliness. I see this as a beginning of a long development in this type of imaging technology," Timlin said.
Potential applications likely will expand as the technology reveals previously unattainable details of cell signaling. Eventually, the Sandia team would li
|Contact: Stephanie Hobby|
DOE/Sandia National Laboratories