Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging

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 reconvolut...

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Bibliographic Details
Published in:Optics express 2011-07, Vol.19 (15), p.13848-13861
Main Authors: Talbot, Clifford B, Patalay, Rakesh, Munro, Ian, Warren, Sean, Ratto, Fulvio, Matteini, Paolo, Pini, Roberto, Breunig, H Georg, König, Karsten, Chu, Antony C, Stamp, Gordon W, Neil, Mark A A, French, Paul M W, Dunsby, Chris
Format: Article
Language:English
Subjects:
Gold - chemistry
Humans
Imaging, Three-Dimensional - instrumentation
Imaging, Three-Dimensional - methods
In Vitro Techniques
Luminescence
Microscopy, Fluorescence, Multiphoton - instrumentation
Microscopy, Fluorescence, Multiphoton - methods
Nanotubes
Spectrometry, Fluorescence
Time Factors
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Summary: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.
ISSN:1094-4087
1094-4087
DOI:10.1364/OE.19.013848