Cellular organelles are subunits that perform specific cellular functions. Recent studies have shown that variations, or heterogeneity, in organelles such as the mitochondria and chloroplasts play an essential role in processes such as molecular metabolism, programmed cell death and energy production. Experimental measurements that study only the average of large populations of organelles are standard, but insufficient for emerging needs. Understanding individual variations requires a new method for studying single organelle physiology with sufficient resolution. The device developed in this project, a platform for self-referencing optrodic sensing, is based on integration of a frequency domain lifetime fluorometer with digital data acquisition and nano- positioning, allowing measurement of chemical fluxes as low as atto mol (10^-18) per square centimeter per second. The instrument will enable exploring the limits of biology in the new realm of organelle physiology. Keystone tests will be the first to measure oxygen flux from single isolated mitochondria or chloroplasts. Coordination with the Purdue Discovery Learning Center will provide broad interdisciplinary opportunities for undergraduate research by emphasizing the interaction between biologists and engineers. Working with the Burton D. Morgan Entrepreneurial Center will facilitate outreach and commercialization of the novel instrumentation and technology.
