Determination of hair cell metabolic state in isolated cochlear preparations by two-photon microscopy

Leann M. Tiede, Sonia Rocha-Sanchez, Richard J. Hallworth, Michael G. Nichols, Kirk Beisel

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Currently there is no accepted method to measure the metabolic status of the organ of Corti. Since metabolism and mito-chondrial dysfunction are expected to play a role in many different hearing disorders, here for the first time we employ two-photon metabolic imaging to assess the metabolic status of the cochlea. When excited with ultrashort pulses of 740-nm light, both inner and outer hair cells in isolated murine cochlear preparations exhibited intrinsic fluorescence. This fluorescence is characterized and shown to be consistent with a mixture of oxidized flavoproteins (Fp) and reduced nicotinamide adenine dinucleotide (NADH). The location of the fluorescence within hair cells is also consistent with the different mitochondrial distributions in these cell types. Treatments with cyanide and mitochondrial uncouplers show that hair cells are metaboli-cally active. Both NADH and Fp in inner hair cells gradually become completely oxidized within 50 min from the time of death of the animal. Outer hair cells show similar trends but are found to have greater variability. We show that it is possible to use two-photon metabolic imaging to assess metabolism in the mouse organ of Corti.

Original languageEnglish
Article number021004
JournalJournal of Biomedical Optics
Volume12
Issue number2
DOIs
StatePublished - Mar 2007

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Outer Auditory Hair Cells
Inner Auditory Hair Cells
Cochlea
hair
Photons
NAD
Organ of Corti
Flavoproteins
Microscopy
Microscopic examination
Fluorescence
Cells
microscopy
preparation
photons
Hearing Disorders
metabolism
Metabolism
organs
fluorescence

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Radiological and Ultrasound Technology
  • Clinical Biochemistry

Cite this

Determination of hair cell metabolic state in isolated cochlear preparations by two-photon microscopy. / Tiede, Leann M.; Rocha-Sanchez, Sonia; Hallworth, Richard J.; Nichols, Michael G.; Beisel, Kirk.

In: Journal of Biomedical Optics, Vol. 12, No. 2, 021004, 03.2007.

Research output: Contribution to journalArticle

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