MICROSCOPY/LIVE CELL IMAGING/CELL BIOLOGY: First STED imaging of live cells in two colors

Sept. 1, 2011
Stimulated emission depletion (STED) microscopy—a super-resolution approach that reveals cells and cellular components in detail by absorbing and releasing energy in fluorescent dye—has been limited to single-color imaging of living cells.

Stimulated emission depletion (STED) microscopy—a super-resolution approach that reveals cells and cellular components in detail by absorbing and releasing energy in fluorescent dye—has been limited to single-color imaging of living cells. But effective study of active cell processes, such as protein interactions, really requires multicolor imaging. Now, a team of researchers from Yale University has given a boost to cell biology by reaching this goal. The group describes its work in the August 2011 issue of Optics Express.1

Tubulin and lamin, immunostained with ATTO 647N and KK 114, respectively; a) Raw intensity STED data, and b) channels decomposed by lifetime separation (green: tubulin, red: lamin). (Images courtesy Optics Express)

The key to the researchers’ achievement was overcoming the challenges of labeling target proteins in living cells with dyes optimal for two-color STED microscopy. By incorporating fusion proteins, they improved the targeting between the protein and the dye, effectively bridging the gap. This enabled them to reach resolutions of 78 and 82 nm for 22 sequential two-color scans of two proteins—epidermal growth factor and epidermal growth factor receptor—in living cells.

The researchers expect that this and other novel approaches will expand live cell STED microscopy to three and more colors, eventually enabling imaging in three dimensions.

1. J. Bückers et al., Opt. Exp. 19, 3130–3143 (2011).

More BioOptics World Current Issue Articles
More BioOptics World Archives Issue Articles

Sponsored Recommendations

Hexapod 6-DOF Active Optical Alignment Micro-Robots - Enablers for Advanced Camera Manufacturing

Dec. 18, 2024
Optics and camera manufacturing benefits from the flexibility of 6-Axis hexapod active optical alignment robots and advanced motion control software

Laser Assisted Wafer Slicing with 3DOF Motion Stages

Dec. 18, 2024
Granite-based high-performance 3-DOF air bearing nanopositioning stages provide ultra-high accuracy and reliability in semiconductor & laser processing applications.

Steering Light: What is the Difference Between 2-Axis Galvo Scanners and Single Mirror 2-Axis Scanners

Dec. 18, 2024
Advantages and limitations of different 2-axis light steering methods: Piezo steering mirrors, voice-coil mirrors, galvos, gimbal mounts, and kinematic mounts.

Free Space Optical Communication

Dec. 18, 2024
Fast Steering Mirrors (FSM) provide fine steering precision to support the Future of Laser Based Communication with LEO Satellites

Voice your opinion!

To join the conversation, and become an exclusive member of Laser Focus World, create an account today!