Developing New Methods to Analyze Cells
When an immune T cell interacts with a target cell, signals must be transmitted through its outer membrane – but how? Dylan Owen and his research team, based at King’s College London, study how two different structures – the T cell’s actin cytoskeleton and outer membrane itself – interact to make this happen.
The structured illumination microscope (SIM) at the AIC is perfectly suited to address this question due to its ability to perform multi-color live cell imaging at twice the resolution offered by conventional microscopes. Using AIC’s high-speed SIM system, Owen’s team could observe the flow of differentially labeled molecules of the actin cytoskeleton and the cell membrane simultaneously. They found that the two molecular flows were correlated with each other, suggesting that the structures are in fact coupled.
Surface T cell receptors, which help the T cell recognize foreign cells to attack, are composed of two protein subunits and several accessory molecules. Back at King’s College, Owen’s team has worked to characterize the T cell receptor structure using localization super-resolution microscopy. To determine whether those molecules were clustered or randomly distributed, Owen’s group developed statistical method to analyze the pointillist 2-dimensional data. This work was published in Nature Methods last year. To further push their statistical approach to studying molecular clusters in 3D biological space, they returned to Janelia to gather data using the AIC’s Interferometric Photo-Activated Localization Microscope (iPALM) – unique in its capability to image individual biomolecules in three dimensions.
Although the team had a very ambitious research goal, “everything worked exactly as we planned,” said Owen. “We wrote a schedule of what we wanted to do on each day, and we got it all done, both times.”
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