On-column labeling for capillary electrophoretic analysis of individual mitochondria directly sampled from tissue cross sections |
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| Authors: | Hossein Ahmadzadeh LaDora V Thompson Edgar A Arriaga |
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| Institution: | (1) Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA;(2) Physical Medicine and Rehabilitation, University of Minnesota, Minneapolis, MN 55455, USA;(3) Present address: Department of Chemistry, California State polytechnic University, Pomona, CA 91768, USA |
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| Abstract: | This technical note reports on a new procedure to on-column-label organelles sampled from a tissue cross section into a fused
silica capillary. These organelles are then analyzed by capillary electrophoresis with postcolumn laser-induced fluorescence
detection. In this procedure, the fluorescent label does not come in contact with the tissue, which facilitates visualization
of the sampled tissue cross section. In addition, on-column labeling allows for better control of the reaction time and fluorescent
label concentrations. As a proof-of-principle, we show results of mitochondria from rat gastrocnemius muscle cross sections
that were on-column-labeled with 10-N-nonyl acridine orange (NAO), a mitochondrion-specific probe, and compare them with results for NAO in-tissue labeling of
the same tissue. The new organelle labeling procedure reported here may easily be extended to the analysis of individual organelles
in other biological samples and may become a valuable tool in studies investigating the role of mitochondria in muscle aging
and exercise physiology.
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| Keywords: | On-column labeling Capillary electrophoresis Fluorescence Mitochondria Muscle |
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