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

Akos A. Gerencser, Judit Doczi, B. Törőcsik, Ella Bossy-Wetzel, V. Ádám-Vizi

Research output: Contribution to journalArticle

39 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

Fingerprint

Mitochondrial Swelling
Thinness
Astrocytes
Neurons
Mitochondria
Fluorescence
Oxidative Phosphorylation
Fluorescence Microscopy
Confocal Microscopy
Proteins

ASJC Scopus subject areas

  • Biophysics

Cite this

Mitochondrial swelling measurement in situ by optimized spatial filtering : Astrocyte-neuron differences. / Gerencser, Akos A.; Doczi, Judit; Törőcsik, B.; Bossy-Wetzel, Ella; Ádám-Vizi, V.

In: Biophysical Journal, Vol. 95, No. 5, 01.09.2008, p. 2583-2598.

Research output: Contribution to journalArticle

@article{08cae8c7991b4242b516b4897c692d64,
title = "Mitochondrial swelling measurement in situ by optimized spatial filtering: Astrocyte-neuron differences",
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.",
author = "Gerencser, {Akos A.} and Judit Doczi and B. T{\"o}rőcsik and Ella Bossy-Wetzel and V. {\'A}d{\'a}m-Vizi",
year = "2008",
month = "9",
day = "1",
doi = "10.1529/biophysj.107.118620",
language = "English",
volume = "95",
pages = "2583--2598",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "5",

}

TY - JOUR

T1 - Mitochondrial swelling measurement in situ by optimized spatial filtering

T2 - Astrocyte-neuron differences

AU - Gerencser, Akos A.

AU - Doczi, Judit

AU - Törőcsik, B.

AU - Bossy-Wetzel, Ella

AU - Ádám-Vizi, V.

PY - 2008/9/1

Y1 - 2008/9/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=51649086458&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=51649086458&partnerID=8YFLogxK

U2 - 10.1529/biophysj.107.118620

DO - 10.1529/biophysj.107.118620

M3 - Article

C2 - 18424491

AN - SCOPUS:51649086458

VL - 95

SP - 2583

EP - 2598

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 5

ER -