Fluorescence Recovery After Photobleaching (FRAP) with simultaneous Fluorescence Lifetime and time-resolved Fluorescence Anisotropy Imaging (FLIM & tr-FAIM)

Y. Teijeiro-Gonzalez, A. Le Marois, A. M. Economou, L. M. Hirvonen, J. A. Levitt, A. J. Beavil, R. L. Beavil, A. Crnjar, C. Molteni, E. Ortiz-Zapater, M. Parsons, K. Suhling

Research output: Chapter in Book/Conference paperConference paper

1 Citation (Scopus)

Abstract

We report the simultaneous combination of three powerful techniques in fluorescence microscopy: Fluorescence Lifetime Imaging (FLIM), Fluorescence Anisotropy Imaging (FAIM) and Fluorescence Recovery After Photobleaching (FRAP), also called F3 microscopy. An exhaustive calibration of the setup was carried out with several rhodamine 6G (R6G) solutions in water-glycerol and from the combination of the FAIM and FRAP data, the hydrodynamic radius of the dye was directly calculated. The F3 data was analyzed with a home-built MATLAB script, and the setup is currently explored further with Green Fluorescent Protein (GFP). Some molecular dynamic (MD) simulations are currently being run in order to help with the interpretation of the experimental anisotropy data.

Original languageEnglish
Title of host publicationThree-Dimensional and Multidimensional Microscopy
Subtitle of host publicationImage Acquisition and Processing XXVI
EditorsTony Wilson, Thomas G. Brown
Place of PublicationUSA
PublisherSPIE-INT SOC OPTICAL ENGINEERING
Volume10883
ISBN (Electronic)9781510624085
DOIs
Publication statusPublished - 21 Feb 2019
Externally publishedYes
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI 2019 - San Francisco, United States
Duration: 5 Feb 20197 Feb 2019

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume10883
ISSN (Print)1605-7422

Conference

ConferenceThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI 2019
CountryUnited States
CitySan Francisco
Period5/02/197/02/19

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