High-throughput capillary-electrophoresis analysis of the contents of a single mitochondria.
We present a technique for labeling the contents of acidic organelles and rapidly releasing, separating, and detecting their labeled contents with laser-induced fluorescence. We have performed solution-phase separation of the contents of single mitochondria and single 100 nm vesicles, which represents a demonstration of an analyzed volume of approximately 1 aL. Our strategy to label the acidic contents of the mitochondrion relies on the use of the membrane-permeable dye, Oregon Green diacetate succinimidyl ester, and a membrane-permeable base to raise intramitochondrial pH. In order to measure the contents, we utilized a glass microfluidic chip and high voltage gradient for millisecond capillary electrophoresis separation after single-mitochondrion photolysis. We observed heterogeneity among a population of mitochondria with respect to a constituent chemical component.
Allen
Peter B
PB
University of Washington, Department of Chemistry, Box 351700, Seattle, Washington 98195, USA.
Doepker
Byron R
BR
Chiu
Daniel T
DT