Luminescence Lifetime Imaging of Chemical Sensors—A Comparison between Time-Domain and Frequency-Domain Based Camera Systems

Koren, Klaus; Moßhammer, Maria; Scholz, Vincent V.; Borisov, Sergey; Holst, Gerhard A.; Kühl, Michael

doi:10.1021/acs.analchem.8b05869

Cited by 39 publications

(31 citation statements)

References 46 publications

(83 reference statements)

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“…Also, this method avoids the use of an additional reference dye common in ratiometric imaging 17,37 . In comparison to other lifetime based camera systems, such as commonly used gated time-domain cameras 8,26 , the novel camera system and protocol presented here can deliver comparable results. In a recent publication, the analytical characteristics of those two systems were compared and it was found that the frequency-domain-based luminescence lifetime camera system is at least comparable to the discontinued time-domain-based predecessor 8 .…”

Section: Discussionmentioning

confidence: 80%

“…The given protocol enables frequency-domain-based luminescence lifetime imaging of the sample in contact with the planar optode using the frequency-domain-based luminescence lifetime camera. More details on this camera system such as the mode of image acquisition and scientific complementary metal-oxide-semiconductor (SCMOS) camera characteristics are given in recent publications 8,29 .…”

Section: Representative Resultsmentioning

confidence: 99%

“…The distribution and dynamics of dissolved gases and ions in sediments and soils provide key information on biogeochemical processes such as microbial respiration 1,2 , or radial oxygen loss from plant roots 3,4,5 , and the chemical microenvironment of microbes 6,7 , plant rhizospheres 5,8,9 and animal burrows 10,11,12 . Biological and chemical activity in such diffusion-limited environments can create steep gradients of chemical substrates or products of biogeochemical processes.…”

Section: Introductionmentioning

confidence: 99%

“…The change between excitation and emission amplitude (i.e., the so-called modulation index or depth (amplitude divided by the constant luminescence part)) is also dependent on the luminescence lifetime. So, by setting a known modulation frequency the special CMOS image sensor within the camera is able to measure the luminescence lifetime in the ns to µs range as described in detail elsewhere 8,29,30 . A general guide on the operation principle can be found (using the following link https://www.youtube.com/watch?v=xPAB_eVWOr8).…”

Section: Introductionmentioning

confidence: 99%

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Luminescence Lifetime Imaging of O<sub>2</sub> with a Frequency-Domain-Based Camera System

Moßhammer

Scholz

Holst³

et al. 2019

JoVE

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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.

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Section: Discussionmentioning

confidence: 80%

Section: Representative Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Luminescence Lifetime Imaging of O<sub>2</sub> with a Frequency-Domain-Based Camera System

Moßhammer

Scholz

Holst³

et al. 2019

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“…1A whereby the phosphorescence of Pt-Glc is specifically quenched by molecular oxygen (O2) and the decay curves collected via various PLIM imaging techniques (e.g. time-domain, frequency modulation or other means [40][41][42] ) are used for calculating phosphorescence lifetimes and subsequent calibrations. PLIM enables improved multiplexed tracing of cell types present in the organoid (crypt and villi regions), e.g.…”

Section: Methods 1: Analysis Of Oxygenation By Phosphorescence Lifetimmentioning

confidence: 99%

A deeper understanding of intestinal organoid metabolism revealed by combining fluorescence lifetime imaging microscopy (FLIM) and extracellular flux analyses

Okkelman

Neto

Papkovsky

et al. 2019

Preprint

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Stem cells and the niche in which they reside feature a complex microenvironment with tightly regulated homeostasis, cell-cell interactions and dynamic regulation of metabolism. A significant number of organoid models has been described over the last decade, yet few methodologies can enable single cell level resolution analysis of the stem cell niche metabolic demands, in real-time and without perturbing integrity. Here, we studied the redox metabolism of Lgr5-GFP intestinal organoids by two emerging microscopy approaches based on luminescence lifetime measurement -fluorescencebased FLIM for NAD(P)H, and phosphorescence-based PLIM for real-time oxygenation. We found that exposure of stem (Lgr5-GFP) and differentiated (no GFP) cells to high and low glucose concentrations resulted in measurable shifts in oxygenation and redox status. NAD(P)H-FLIM and O2-PLIM both indicated that at high 'basal' glucose conditions, Lgr5-GFP cells had lower activity of oxidative phosphorylation when compared with cells lacking Lgr5. However, when exposed to low (0.5 mM) glucose, stem cells utilized oxidative metabolism more dynamically than non-stem cells. The high heterogeneity of complex 3D architecture and energy production pathways of Lgr5-GFP organoids were also confirmed by the extracellular flux (XF) analysis. Our data reveals that combined analysis of NAD(P)H-FLIM and organoid oxygenation by PLIM represents promising approach for studying stem cell niche metabolism in a live readout.

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Kabelbakterienskelette als katalytisch aktive Elektroden

Digel,

Mierzwa,

Bonné

et al. 2023

Angewandte Chemie

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Kabelbakterien sind vielzellige, filamentöse Bakterien, die interne leitfähige Fasern verwenden, um Elektronen über Zentimeterdistanzen von Elektronenspendern in anoxischen Sedimentschichten zu Sauerstoff an der Oberfläche zu transportieren. Wir haben diese Fasern extrahiert und sie als freistehende biobasierte Elektroden verwendet, um ihr elektrokatalytisches Verhalten zu untersuchen. Die Fasern katalysieren die reversible Umwandlung von Sauerstoff und Wasser, kombiniert mit einem elektrischen Stromfluss durch die Fasern, selbst wenn die Potentialdifferenz ausschließlich durch einen Sauerstoffkonzentrationsgradienten erzeugt wird. Sowohl die Sauerstoffreduktion als auch die Sauerstoffentwicklung wurden durch optische Messungen bestätigt. Bei lebenden Kabelbakterien erklärt sich die Sauerstoffreduktion durch direkte Elektrokatalyse an den Fasern, und nicht durch membrangebundene Proteine, und führt zu den außergewöhnlich hohen zellspezifischen Sauerstoffverbrauchsraten, die in der oxischen Zone beobachtet werden, während die elektrokatalytische Wasseroxidation Zellen in der anoxischen Zone mit Sauerstoff versorgen kann.

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Luminescence Lifetime Imaging of Chemical Sensors—A Comparison between Time-Domain and Frequency-Domain Based Camera Systems

Cited by 39 publications

References 46 publications

Luminescence Lifetime Imaging of O<sub>2</sub> with a Frequency-Domain-Based Camera System

Luminescence Lifetime Imaging of O<sub>2</sub> with a Frequency-Domain-Based Camera System

A deeper understanding of intestinal organoid metabolism revealed by combining fluorescence lifetime imaging microscopy (FLIM) and extracellular flux analyses

Kabelbakterienskelette als katalytisch aktive Elektroden

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