Luminescence Lifetime Imaging of O2 with a Frequency-Domain-Based Camera System

Maria Moßhammer, Vincent V. Scholz, Gerhard Holst, Michael Kühl, Klaus Koren

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Abstract

We describe a method to image dissolved oxygen (O 2), in 2D at high spatial (< 50-100 µm) and temporal (< 10 s) resolution. The method employs O 2 sensitive luminescent sensor foils (planar optodes) in combination with a specialized camera system for imaging luminescence lifetime in the frequency-domain. Planar optodes are prepared by dissolving the O 2-sensitive indicator dye in a polymer and spreading the mixture on a solid support in a defined thickness via knife coating. After evaporation of the solvent, the planar optode is placed in close contact with the sample of interest-here demonstrated with the roots of the aquatic plant Littorella uniflora. The O 2 concentration-dependent change in the luminescence lifetime of the indicator dye within the planar optode is imaged via the backside of the transparent carrier foil and aquarium wall using a special camera. This camera measures the luminescence lifetime (µs) via a shift in phase angle between a modulated excitation signal and emission signal. This method is superior to luminescence intensity imaging methods, as the signal is independent of the dye concentration or intensity of the excitation source, and solely relies on the luminescence decay time, which is an intrinsically referenced parameter. Consequently, an additional reference dye or other means of referencing are not needed. We demonstrate the use of the system for macroscopic O 2 imaging of plant rhizospheres, but the camera system can also easily be coupled to a microscope.

OriginalsprogEngelsk
Artikelnummere60191
TidsskriftJournal of visualized experiments : JoVE
Vol/bind2019
Nummer154
ISSN1940-087X
DOI
StatusUdgivet - dec. 2019

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