Dataset Information

Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions.

ABSTRACT: Recent evidence suggests that the activity of mitochondrial oxidative phosphorylation complexes (MOPCs) is modulated at multiple sites. Here, a method of optically monitoring electron distribution within and between MOPCs is described using a center-mounted sample in an integrating sphere (to minimize scattering effects) with a rapid-scanning spectrometer. The redox-sensitive MOPC absorbances (∼465-630 nm) were modeled using linear least squares analysis with individual chromophore spectra. Classical mitochondrial activity transitions (e.g., ADP-induced increase in oxygen consumption) were used to characterize this approach. Most notable in these studies was the observation that intermediates of the catalytic cycle of cytochrome oxidase are dynamically modulated with metabolic state. The MOPC redox state, along with measurements of oxygen consumption and mitochondrial membrane potential, was used to evaluate the conductances of different sections of the electron transport chain. This analysis then was applied to mitochondria isolated from rabbit hearts subjected to ischemia/reperfusion (I/R). Surprisingly, I/R resulted in an inhibition of all measured MOPC conductances, suggesting a coordinated down-regulation of mitochondrial activity with this well-established cardiac perturbation.

SUBMITTER: Chess DJ

PROVIDER: S-EPMC3713082 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions.

Chess David J DJ Billings Eric E Covian Raúl R Glancy Brian B French Stephanie S Taylor Joni J de Bari Heather H Murphy Elizabeth E Balaban Robert S RS

Analytical biochemistry 20130509 2

Recent evidence suggests that the activity of mitochondrial oxidative phosphorylation complexes (MOPCs) is modulated at multiple sites. Here, a method of optically monitoring electron distribution within and between MOPCs is described using a center-mounted sample in an integrating sphere (to minimize scattering effects) with a rapid-scanning spectrometer. The redox-sensitive MOPC absorbances (∼465-630 nm) were modeled using linear least squares analysis with individual chromophore spectra. Clas ...[more]

PMID: 23665273

Similar Datasets

Project description:We introduce a multi-distance, frequency-domain, near-infrared spectroscopy (NIRS) method to measure the optical coefficients of two-layered media and the thickness of the top layer from diffuse reflectance measurements. This method features a direct solution based on diffusion theory and an inversion procedure based on the Levenberg-Marquardt algorithm. We have validated our method through Monte Carlo simulations, experiments on tissue-like phantoms, and measurements on the forehead of three human subjects. The Monte Carlo simulations and phantom measurements have shown that, in ideal two-layered samples, our method accurately recovers the top layer thickness (L), the absorption coefficient (µ a ) and the reduced scattering coefficient (µ' s ) of both layers with deviations that are typically less than 10% for all parameters. Our method is aimed at absolute measurements of hemoglobin concentration and saturation in cerebral and extracerebral tissue of adult human subjects, where the top layer (layer 1) represents extracerebral tissue (scalp, skull, dura mater, subarachnoid space, etc.) and the bottom layer (layer 2) represents cerebral tissue. Human subject measurements have shown a significantly greater total hemoglobin concentration in cerebral tissue (82±14 µM) with respect to extracerebral tissue (30±7 µM). By contrast, there was no significant difference between the hemoglobin saturation measured in cerebral tissue (56%±10%) and extracerebral tissue (62%±6%). To our knowledge, this is the first time that an inversion procedure in the frequency domain with six unknown parameters with no other prior knowledge is used for the retrieval of the optical coefficients and top layer thickness with high accuracy on two-layered media. Our absolute measurements of cerebral hemoglobin concentration and saturation are based on the discrimination of extracerebral and cerebral tissue layers, and they can enhance the impact of NIRS for cerebral hemodynamics and oxygenation assessment both in the research arena and clinical practice.

Dataset Information

Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions.

Publications

Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets