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Skeletal Muscle NAD(P)H Two-Photon Fluorescence Microscopy In Vivo: Topology and Optical Inner Filters

ABSTRACT

Two-photon excitation fluorescence microscopy (TPEFM) permits the investigation of the topology of intercellular events within living animals. TPEFM was used to monitor the distribution of mitochondrial reduced nicotinamide adenine dinucleotide (NAD(P)H) in murine skeletal muscle in vivo. NAD(P)H fluorescence emission was monitored (~460 nm) using 710-720 nm excitation. High-resolution TPEFM images were collected up to a depth of 150 µm from the surface of the tibialis anterior muscle. The NAD(P)H fluorescence images revealed subcellular structures consistent with subsarcolemmal, perivascular, intersarcomeric, and paranuclear mitochondria. In vivo fiber typing between IIB and ...

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