TY - JOUR
T1 - Nanohole Array-Directed Trapping of Mammalian Mitochondria Enabling Single Organelle Analysis
AU - Kumar, Shailabh
AU - Wolken, Gregory G.
AU - Wittenberg, Nathan J.
AU - Arriaga, Edgar A.
AU - Oh, Sang Hyun
N1 - Publisher Copyright:
© 2015 American Chemical Society.
PY - 2015/12/15
Y1 - 2015/12/15
N2 - We present periodic nanohole arrays fabricated in free-standing metal-coated nitride films as a platform for trapping and analyzing single organelles. When a microliter-scale droplet containing mitochondria is dispensed above the nanohole array, the combination of evaporation and capillary flow directs individual mitochondria to the nanoholes. Mammalian mitochondria arrays were rapidly formed on chip using this technique without any surface modification steps, microfluidic interconnects, or external power sources. The trapped mitochondria were depolarized on chip using an ionophore with results showing that the organelle viability and behavior were preserved during the on-chip assembly process. Fluorescence signal related to mitochondrial membrane potential was obtained from single mitochondria trapped in individual nanoholes revealing statistical differences between the behavior of polarized vs depolarized mammalian mitochondria. This technique provides a fast and stable route for droplet-based directed localization of organelles-on-a-chip with minimal limitations and complexity, as well as promotes integration with other optical or electrochemical detection techniques.
AB - We present periodic nanohole arrays fabricated in free-standing metal-coated nitride films as a platform for trapping and analyzing single organelles. When a microliter-scale droplet containing mitochondria is dispensed above the nanohole array, the combination of evaporation and capillary flow directs individual mitochondria to the nanoholes. Mammalian mitochondria arrays were rapidly formed on chip using this technique without any surface modification steps, microfluidic interconnects, or external power sources. The trapped mitochondria were depolarized on chip using an ionophore with results showing that the organelle viability and behavior were preserved during the on-chip assembly process. Fluorescence signal related to mitochondrial membrane potential was obtained from single mitochondria trapped in individual nanoholes revealing statistical differences between the behavior of polarized vs depolarized mammalian mitochondria. This technique provides a fast and stable route for droplet-based directed localization of organelles-on-a-chip with minimal limitations and complexity, as well as promotes integration with other optical or electrochemical detection techniques.
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U2 - 10.1021/acs.analchem.5b03604
DO - 10.1021/acs.analchem.5b03604
M3 - Article
C2 - 26593329
AN - SCOPUS:84950136566
SN - 0003-2700
VL - 87
SP - 11973
EP - 11977
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 24
ER -