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

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.

SUBMITTER: Allen PB

PROVIDER: S-EPMC2682631 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature

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

Allen Peter B PB Doepker Byron R BR Chiu Daniel T DT

Analytical chemistry 20090501 10

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 s ...[more]

PMID: 19344146

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