The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion

Peter H.G.M. Willems; Herman G. Swarts; Mark A. Hink; Werner J.H. Koopman

doi:10.1016/S0076-6879(08)04416-9

The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion

Peter H.G.M. Willems^*, Herman G. Swarts, Mark A. Hink, Werner J.H. Koopman

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

8 Citations (Scopus)

Abstract

Within cells, functional changes in mitochondrial metabolic state are associated with alterations in organelle mobility, shape, and configuration of the mitochondrial matrix. Fluorescence correlation spectroscopy (FCS) is a technique that measures intensity fluctuations caused by single fluorescent molecules moving through a small detection volume. By mathematically correlating these fluctuations, information can be obtained about the concentration and rate of diffusion of the fluorescent molecules. Here we present an FCS-based approach for determining the mobility of enhanced yellow fluorescent protein (mitoEYFP) in the mitochondrial matrix of primary human skin fibroblasts. This protocol allows simultaneous quantification of intramatrix EYFP concentration and its diffusion constant, as well as the fraction of moving mitochondria and their velocity.

Original language	English
Title of host publication	Mitochondrial Function, Part A
Subtitle of host publication	Mitochondrial Electron Transport Complexes and Reactive Oxygen Species
Editors	William Allison, Immo Scheffler
Publisher	Elsevier
Chapter	16
Pages	287-302
Number of pages	16
Edition	A
ISBN (Print)	9780080877761
DOIs	https://doi.org/10.1016/S0076-6879(08)04416-9
Publication status	Published - 2009
Externally published	Yes

Publication series

Name	Methods in Enzymology
Number	A
Volume	456
ISSN (Print)	0076-6879

Access to Document

10.1016/S0076-6879(08)04416-9

8 Citations
1 Chapter

Tracing Human Mitochondrial Complex I Assembly by Use of GFP-Tagged Subunits
Dieteren, C. E. J., Koopman, W. J. H. & Nijtmans, L. G. J., 2009, Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. Allison, W. & Scheffler, I. (eds.). Elsevier, p. 133-151 19 p. (Methods in Enzymology; vol. 456, no. A).
Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review
6 Citations (Scopus)

Cite this

Willems, P. H. G. M., Swarts, H. G., Hink, M. A., & Koopman, W. J. H. (2009). The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion. In W. Allison, & I. Scheffler (Eds.), Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species (A ed., pp. 287-302). (Methods in Enzymology; Vol. 456, No. A). Elsevier. https://doi.org/10.1016/S0076-6879(08)04416-9

Willems, Peter H.G.M. ; Swarts, Herman G. ; Hink, Mark A. et al. / The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion. Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. editor / William Allison ; Immo Scheffler. A. ed. Elsevier, 2009. pp. 287-302 (Methods in Enzymology; A).

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abstract = "Within cells, functional changes in mitochondrial metabolic state are associated with alterations in organelle mobility, shape, and configuration of the mitochondrial matrix. Fluorescence correlation spectroscopy (FCS) is a technique that measures intensity fluctuations caused by single fluorescent molecules moving through a small detection volume. By mathematically correlating these fluctuations, information can be obtained about the concentration and rate of diffusion of the fluorescent molecules. Here we present an FCS-based approach for determining the mobility of enhanced yellow fluorescent protein (mitoEYFP) in the mitochondrial matrix of primary human skin fibroblasts. This protocol allows simultaneous quantification of intramatrix EYFP concentration and its diffusion constant, as well as the fraction of moving mitochondria and their velocity.",

author = "Willems, {Peter H.G.M.} and Swarts, {Herman G.} and Hink, {Mark A.} and Koopman, {Werner J.H.}",

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language = "English",

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Willems, PHGM, Swarts, HG, Hink, MA & Koopman, WJH 2009, The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion. in W Allison & I Scheffler (eds), Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. A edn, Methods in Enzymology, no. A, vol. 456, Elsevier, pp. 287-302. https://doi.org/10.1016/S0076-6879(08)04416-9

The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion. / Willems, Peter H.G.M.; Swarts, Herman G.; Hink, Mark A. et al.
Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. ed. / William Allison; Immo Scheffler. A. ed. Elsevier, 2009. p. 287-302 (Methods in Enzymology; Vol. 456, No. A).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

TY - CHAP

T1 - The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion

AU - Willems, Peter H.G.M.

AU - Swarts, Herman G.

AU - Hink, Mark A.

AU - Koopman, Werner J.H.

PY - 2009

Y1 - 2009

N2 - Within cells, functional changes in mitochondrial metabolic state are associated with alterations in organelle mobility, shape, and configuration of the mitochondrial matrix. Fluorescence correlation spectroscopy (FCS) is a technique that measures intensity fluctuations caused by single fluorescent molecules moving through a small detection volume. By mathematically correlating these fluctuations, information can be obtained about the concentration and rate of diffusion of the fluorescent molecules. Here we present an FCS-based approach for determining the mobility of enhanced yellow fluorescent protein (mitoEYFP) in the mitochondrial matrix of primary human skin fibroblasts. This protocol allows simultaneous quantification of intramatrix EYFP concentration and its diffusion constant, as well as the fraction of moving mitochondria and their velocity.

AB - Within cells, functional changes in mitochondrial metabolic state are associated with alterations in organelle mobility, shape, and configuration of the mitochondrial matrix. Fluorescence correlation spectroscopy (FCS) is a technique that measures intensity fluctuations caused by single fluorescent molecules moving through a small detection volume. By mathematically correlating these fluctuations, information can be obtained about the concentration and rate of diffusion of the fluorescent molecules. Here we present an FCS-based approach for determining the mobility of enhanced yellow fluorescent protein (mitoEYFP) in the mitochondrial matrix of primary human skin fibroblasts. This protocol allows simultaneous quantification of intramatrix EYFP concentration and its diffusion constant, as well as the fraction of moving mitochondria and their velocity.

U2 - 10.1016/S0076-6879(08)04416-9

DO - 10.1016/S0076-6879(08)04416-9

M3 - Chapter

C2 - 19348895

AN - SCOPUS:63449108900

SN - 9780080877761

T3 - Methods in Enzymology

SP - 287

EP - 302

BT - Mitochondrial Function, Part A

A2 - Allison, William

A2 - Scheffler, Immo

PB - Elsevier

ER -

Willems PHGM, Swarts HG, Hink MA, Koopman WJH. The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion. In Allison W, Scheffler I, editors, Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. A ed. Elsevier. 2009. p. 287-302. (Methods in Enzymology; A). doi: 10.1016/S0076-6879(08)04416-9

The Use of Fluorescence Correlation Spectroscopy to Probe Mitochondrial Mobility and Intramatrix Protein Diffusion

Abstract

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Research output

Tracing Human Mitochondrial Complex I Assembly by Use of GFP-Tagged Subunits

Cite this