"This experiment was a proof of the concept," says Liang Chen. "We demonstrated the dynamics of protein phosphorylation in controlling differentiation in this biological system using synchrotron infrared spectromicroscopy, and we pointed the way to answering the many questions a biologist has to ask about measuring the coordination of specific processes in real time."
Although in this first experiment the team was not able to follow individual cells continuously, they were able to monitor differentiation in groups of cultured PC12 cells in real time, without labeling or any other invasive procedure. It was the first step in an ambitious inquiry into the real-time biochemistry of living mammalian cells over the long term.
At beamline 1.4.3., with the help of new team members Kevin Loutherback and Rafael Gomez-Sjoberg, the team is designing equipment to maintain mammalian cells in a thin layer of culture media that will keep them healthy yet not interfere with the infrared beam, while automatically monitoring and adjusting temperature, humidity, and nutrient ratios, and removing waste products. This will allow data on individual cells to be gathered continuously throughout the entire phosphorylation process.
Meanwhile the Berkeley Synchrotron Infrared Structural Biology program at ALS beamline 5.4 is building multimodal facilities that will monitor cell development in human cells, bacteria, and plants, within soils, minerals, and other environments, via "hyperspectromicroscopy" from the ultraviolet through visible light and deep into the infrared. Researchers will be able to choose the frequency window (or combination of windows) best suited to the sample and the conditions in Holman's words, "to watch almost everything at once."
Says Holman, "Many researchers from the medical communities are interested in using the technology, and we are particularly interested in collaborating with university centers and private firm
|Contact: Paul Preuss|
DOE/Lawrence Berkeley National Laboratory