Fluorescence lifetime imaging for viscosity and diffusion measurements

Klaus Suhling; Yurema Teijeiro-Gonzalez; I. Emilie Steinmark; Arjuna L. James; Augoustina M. Economou; Gokhan Yahioglu; Alix Le Marois; Liisa M. Hirvonen; Jakub Nedbal; James A. Levitt; Pei Hua Chung; Cécile A. Dreiss; Andrew J. Beavil; Rebecca L. Beavil; Elena Ortiz-Zapater; Christian D. Lorenz; Maddy Parsons; Alessandro Crnjar; Bethan Cornell; Carla Molteni

doi:10.1117/12.2508744

Fluorescence lifetime imaging for viscosity and diffusion measurements

Klaus Suhling, Yurema Teijeiro-Gonzalez, I. Emilie Steinmark, Arjuna L. James, Augoustina M. Economou, Gokhan Yahioglu, Alix Le Marois, Liisa M. Hirvonen, Jakub Nedbal, James A. Levitt, Pei Hua Chung, Cécile A. Dreiss, Andrew J. Beavil, Rebecca L. Beavil, Elena Ortiz-Zapater, Christian D. Lorenz, Maddy Parsons, Alessandro Crnjar, Bethan Cornell, Carla Molteni

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

4 Citations (Scopus)

Abstract

Imaging viscosity and its spatiotemporal patterns can provide valuable insight into the underlying physical conditions of biochemical reactions and biological processes in cells and tissues. One way to measure viscosity and diffusion is the use of fluorescence recovery after photobleaching (FRAP). We combine FRAP with FLIM and time-resolved fluorescence anisotropy imaging (tr-FAIM), by acquiring time- and polarization-resolved fluorescence images in every frame of a FRAP series. This allows us to simultaneously monitor translational and rotational diffusion. This approach can be applied to measuring diffusion in homogeneous and heterogeneous environments, and in principle also allows the study of homo-FRET. Another way to measure viscosity and diffusion is through specific flexible dyes, e.g. fluorescent molecular rotors, whose fluorescence quantum yield and fluorescence lifetime depend on the viscosity of the environment, in combination with fluorescence lifetime imaging (FLIM). We show that a bodipybased fluorescent molecular rotor targeting mitochondria reports on their viscosity, which changes under physiological stimuli. Both methods can optically measure viscosity and diffusion on the micrometer scale.

Original language	English
Title of host publication	Multiphoton Microscopy in the Biomedical Sciences XIX
Editors	Ammasi Periasamy, Peter T. C. So, Karsten Konig
Publisher	SPIE
Volume	10882
ISBN (Electronic)	9781510624061
DOIs	https://doi.org/10.1117/12.2508744
Publication status	Published - 22 Feb 2019
Event	Multiphoton Microscopy in the Biomedical Sciences XIX 2019 - San Francisco, United States Duration: 3 Feb 2019 → 6 Feb 2019

Conference

Conference	Multiphoton Microscopy in the Biomedical Sciences XIX 2019
Country/Territory	United States
City	San Francisco
Period	3/02/2019 → 6/02/2019

Keywords

Fluorescence lifetime imaging
Fluorescence recovery after photobleaching
Fluorescent molecular rotors
Time-correlated single photon counting
Time-resolved fluorescence anisotropy imaging

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10.1117/12.2508744

Cite this

Suhling, K., Teijeiro-Gonzalez, Y., Steinmark, I. E., James, A. L., Economou, A. M., Yahioglu, G., Le Marois, A., Hirvonen, L. M., Nedbal, J., Levitt, J. A., Chung, P. H., Dreiss, C. A., Beavil, A. J., Beavil, R. L., Ortiz-Zapater, E., Lorenz, C. D., Parsons, M., Crnjar, A., Cornell, B., & Molteni, C. (2019). Fluorescence lifetime imaging for viscosity and diffusion measurements. In A. Periasamy, P. T. C. So, & K. Konig (Eds.), Multiphoton Microscopy in the Biomedical Sciences XIX (Vol. 10882). Article 1088210 SPIE. https://doi.org/10.1117/12.2508744

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title = "Fluorescence lifetime imaging for viscosity and diffusion measurements",

abstract = "Imaging viscosity and its spatiotemporal patterns can provide valuable insight into the underlying physical conditions of biochemical reactions and biological processes in cells and tissues. One way to measure viscosity and diffusion is the use of fluorescence recovery after photobleaching (FRAP). We combine FRAP with FLIM and time-resolved fluorescence anisotropy imaging (tr-FAIM), by acquiring time- and polarization-resolved fluorescence images in every frame of a FRAP series. This allows us to simultaneously monitor translational and rotational diffusion. This approach can be applied to measuring diffusion in homogeneous and heterogeneous environments, and in principle also allows the study of homo-FRET. Another way to measure viscosity and diffusion is through specific flexible dyes, e.g. fluorescent molecular rotors, whose fluorescence quantum yield and fluorescence lifetime depend on the viscosity of the environment, in combination with fluorescence lifetime imaging (FLIM). We show that a bodipybased fluorescent molecular rotor targeting mitochondria reports on their viscosity, which changes under physiological stimuli. Both methods can optically measure viscosity and diffusion on the micrometer scale.",

keywords = "Fluorescence lifetime imaging, Fluorescence recovery after photobleaching, Fluorescent molecular rotors, Time-correlated single photon counting, Time-resolved fluorescence anisotropy imaging",

author = "Klaus Suhling and Yurema Teijeiro-Gonzalez and Steinmark, {I. Emilie} and James, {Arjuna L.} and Economou, {Augoustina M.} and Gokhan Yahioglu and {Le Marois}, Alix and Hirvonen, {Liisa M.} and Jakub Nedbal and Levitt, {James A.} and Chung, {Pei Hua} and Dreiss, {C{\'e}cile A.} and Beavil, {Andrew J.} and Beavil, {Rebecca L.} and Elena Ortiz-Zapater and Lorenz, {Christian D.} and Maddy Parsons and Alessandro Crnjar and Bethan Cornell and Carla Molteni",

year = "2019",

month = feb,

day = "22",

doi = "10.1117/12.2508744",

language = "English",

volume = "10882",

editor = "Ammasi Periasamy and So, {Peter T. C.} and Karsten Konig",

booktitle = "Multiphoton Microscopy in the Biomedical Sciences XIX",

publisher = "SPIE",

address = "United States",

note = "Multiphoton Microscopy in the Biomedical Sciences XIX 2019 ; Conference date: 03-02-2019 Through 06-02-2019",

}

Suhling, K , Teijeiro-Gonzalez, Y , Steinmark, IE, James, AL, Economou, AM, Yahioglu, G, Le Marois, A , Hirvonen, LM , Nedbal, J , Levitt, JA , Chung, PH , Dreiss, CA , Beavil, AJ , Beavil, RL , Ortiz-Zapater, E , Lorenz, CD , Parsons, M , Crnjar, A , Cornell, B & Molteni, C 2019, Fluorescence lifetime imaging for viscosity and diffusion measurements. in A Periasamy, PTC So & K Konig (eds), Multiphoton Microscopy in the Biomedical Sciences XIX. vol. 10882, 1088210, SPIE, Multiphoton Microscopy in the Biomedical Sciences XIX 2019, San Francisco, United States, 3/02/2019. https://doi.org/10.1117/12.2508744

Fluorescence lifetime imaging for viscosity and diffusion measurements. / Suhling, Klaus ; Teijeiro-Gonzalez, Yurema ; Steinmark, I. Emilie et al.
Multiphoton Microscopy in the Biomedical Sciences XIX. ed. / Ammasi Periasamy; Peter T. C. So; Karsten Konig. Vol. 10882 SPIE, 2019. 1088210.

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

TY - CHAP

T1 - Fluorescence lifetime imaging for viscosity and diffusion measurements

AU - Suhling, Klaus

AU - Teijeiro-Gonzalez, Yurema

AU - Steinmark, I. Emilie

AU - James, Arjuna L.

AU - Economou, Augoustina M.

AU - Yahioglu, Gokhan

AU - Le Marois, Alix

AU - Hirvonen, Liisa M.

AU - Nedbal, Jakub

AU - Levitt, James A.

AU - Chung, Pei Hua

AU - Dreiss, Cécile A.

AU - Beavil, Andrew J.

AU - Beavil, Rebecca L.

AU - Ortiz-Zapater, Elena

AU - Lorenz, Christian D.

AU - Parsons, Maddy

AU - Crnjar, Alessandro

AU - Cornell, Bethan

AU - Molteni, Carla

PY - 2019/2/22

Y1 - 2019/2/22

N2 - Imaging viscosity and its spatiotemporal patterns can provide valuable insight into the underlying physical conditions of biochemical reactions and biological processes in cells and tissues. One way to measure viscosity and diffusion is the use of fluorescence recovery after photobleaching (FRAP). We combine FRAP with FLIM and time-resolved fluorescence anisotropy imaging (tr-FAIM), by acquiring time- and polarization-resolved fluorescence images in every frame of a FRAP series. This allows us to simultaneously monitor translational and rotational diffusion. This approach can be applied to measuring diffusion in homogeneous and heterogeneous environments, and in principle also allows the study of homo-FRET. Another way to measure viscosity and diffusion is through specific flexible dyes, e.g. fluorescent molecular rotors, whose fluorescence quantum yield and fluorescence lifetime depend on the viscosity of the environment, in combination with fluorescence lifetime imaging (FLIM). We show that a bodipybased fluorescent molecular rotor targeting mitochondria reports on their viscosity, which changes under physiological stimuli. Both methods can optically measure viscosity and diffusion on the micrometer scale.

AB - Imaging viscosity and its spatiotemporal patterns can provide valuable insight into the underlying physical conditions of biochemical reactions and biological processes in cells and tissues. One way to measure viscosity and diffusion is the use of fluorescence recovery after photobleaching (FRAP). We combine FRAP with FLIM and time-resolved fluorescence anisotropy imaging (tr-FAIM), by acquiring time- and polarization-resolved fluorescence images in every frame of a FRAP series. This allows us to simultaneously monitor translational and rotational diffusion. This approach can be applied to measuring diffusion in homogeneous and heterogeneous environments, and in principle also allows the study of homo-FRET. Another way to measure viscosity and diffusion is through specific flexible dyes, e.g. fluorescent molecular rotors, whose fluorescence quantum yield and fluorescence lifetime depend on the viscosity of the environment, in combination with fluorescence lifetime imaging (FLIM). We show that a bodipybased fluorescent molecular rotor targeting mitochondria reports on their viscosity, which changes under physiological stimuli. Both methods can optically measure viscosity and diffusion on the micrometer scale.

KW - Fluorescence lifetime imaging

KW - Fluorescence recovery after photobleaching

KW - Fluorescent molecular rotors

KW - Time-correlated single photon counting

KW - Time-resolved fluorescence anisotropy imaging

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U2 - 10.1117/12.2508744

DO - 10.1117/12.2508744

M3 - Conference paper

AN - SCOPUS:85067816147

VL - 10882

BT - Multiphoton Microscopy in the Biomedical Sciences XIX

A2 - Periasamy, Ammasi

A2 - So, Peter T. C.

A2 - Konig, Karsten

PB - SPIE

T2 - Multiphoton Microscopy in the Biomedical Sciences XIX 2019

Y2 - 3 February 2019 through 6 February 2019

ER -

Fluorescence lifetime imaging for viscosity and diffusion measurements

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Keywords

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