High-throughput capillary-electrophoresis analysis of the contents of a single mitochondria

Anal Chem. 2009 May 15;81(10):3784-91. doi: 10.1021/ac900099y.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Carboxylic Acids / chemistry
  • Electrophoresis, Capillary / methods*
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Mitochondria / chemistry*

Substances

  • Carboxylic Acids
  • Oregon Green 488 carboxylic acid