title: Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging.
abstract: When performing multiphoton fluorescence lifetime imaging in multiple spectral emission channels, an instrument response function must be acquired in each channel if accurate measurements of complex fluorescence decays are to be performed. Although this can be achieved using the reference reconvolution technique, it is difficult to identify suitable fluorophores with a mono-exponential fluorescence decay across a broad emission spectrum. We present a solution to this problem by measuring the IRF using the ultrafast luminescence from gold nanorods. We show that ultrafast gold nanorod luminescence allows the IRF to be directly obtained in multiple spectral channels simultaneously across a wide spectral range. We validate this approach by presenting an analysis of multispectral autofluorescence FLIM data obtained from human skin ex vivo.
authors.affiliation: Department of Physics, Imperial College London, UK. clifford.talbot@imperial.ac.uk
authors.forename: Clifford B
authors.initials: CB
authors.lastname: Talbot
authors.forename: Rakesh
authors.initials: R
authors.lastname: Patalay
authors.forename: Ian
authors.initials: I
authors.lastname: Munro
authors.forename: Sean
authors.initials: S
authors.lastname: Warren
authors.forename: Fulvio
authors.initials: F
authors.lastname: Ratto
authors.forename: Paolo
authors.initials: P
authors.lastname: Matteini
authors.forename: Roberto
authors.initials: R
authors.lastname: Pini
authors.forename: H Georg
authors.initials: HG
authors.lastname: Breunig
authors.forename: Karsten
authors.initials: K
authors.lastname: König
authors.forename: Antony C
authors.initials: AC
authors.lastname: Chu
authors.forename: Gordon W
authors.initials: GW
authors.lastname: Stamp
authors.forename: Mark A A
authors.initials: MA
authors.lastname: Neil
authors.forename: Paul M W
authors.initials: PM
authors.lastname: French
authors.forename: Chris
authors.initials: C
authors.lastname: Dunsby
mesh_terms: D006046:Gold / Q000737:chemistry*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods*; D066298:In Vitro Techniques; D049449:Luminescence; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods*; D043942:Nanotubes; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D006046:Gold