Mitochondrial swelling measurement in situ by optimized spatial filtering: Astrocyte-neuron differences

Akos A. Gerencser, Judit Doczi, Beata Töröcsik, Ella Bossy-Wetzel, Vera Adam-Vizi

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Mitochondrial swelling is a hallmark of mitochondrial dysfunction, and is an indicator of the opening of the mitochondrial permeability transition pore. We introduce here a novel quantitative in situ single-cell assay of mitochondrial swelling based on standard wide-field or confocal fluorescence microscopy. This morphometric technique quantifies the relative diameter of mitochondria labeled by targeted fluorescent proteins. Fluorescence micrographs are spatial bandpass filtered transmitting either high or low spatial frequencies. Mitochondrial swelling is measured by the fluorescence intensity ratio of the high- to low-frequency filtered copy of the same image. We have termed this fraction the "thinness ratio". The filters are designed by numeric optimization for sensitivity. We characterized the thinness ratio technique by modeling microscopic image formation and by experimentation in cultured cortical neurons and astrocytes. The frequency domain image processing endows robustness and subresolution sensitivity to the thinness ratio technique, overcoming the limitations of shape measurement approaches. The thinness ratio proved to be highly sensitive to mitochondrial swelling, but insensitive to fission or fusion of mitochondria. We found that in situ astrocytic mitochondria swell upon short-term uncoupling or inhibition of oxidative phosphorylation, whereas such responses are absent in cultured cortical neurons.

Original languageEnglish
Pages (from-to)2583-2598
Number of pages16
JournalBiophysical journal
Volume95
Issue number5
DOIs
Publication statusPublished - Sep 1 2008

ASJC Scopus subject areas

  • Biophysics

Fingerprint Dive into the research topics of 'Mitochondrial swelling measurement in situ by optimized spatial filtering: Astrocyte-neuron differences'. Together they form a unique fingerprint.

  • Cite this