Investigation of copper homeostasis in plant cells by fluorescence lifetime imaging microscopy

Hötzer, Benjamin; Ivanov, Rumen; Bauer, Petra; Jung, Gregor

doi:10.4161/psb.19561

Cited by 7 publications

(5 citation statements)

References 23 publications

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“…In biomedicine, fluorescence lifetime measurements and fluorescence lifetime imaging (FLIM) are used to study changes in tissue autofluorescence [ 28 ]. However, fluorescence lifetime techniques have not been widely utilized for plant studies although FLIM has been used to study photosynthesis [ 26 , 29 , 30 ], and the uptake of minerals [ 31 , 32 ] and was previously used to study the effect of a photosynthetic inhibitor DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea) in Chlamydomonas reinhardtii [ 29 ]. We report here the application of fluorescence lifetime imaging of chlorophyll to provide a label-free in vivo means to non-invasively map the effect of a PS II inhibiting herbicide in Triticum aestivum (Winter Wheat) on different spatial scales through its local impact on chlorophyll fluorescence.…”

Section: Introductionmentioning

confidence: 99%

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

et al. 2017

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BackgroundIn order to better understand and improve the mode of action of agrochemicals, it is useful to be able to visualize their uptake and distribution in vivo, non-invasively and, ideally, in the field. Here we explore the potential of plant autofluorescence (specifically chlorophyll fluorescence) to provide a readout of herbicide action across the scales utilising multiphoton-excited fluorescence lifetime imaging, wide-field single-photon excited fluorescence lifetime imaging and single point fluorescence lifetime measurements via a fibre-optic probe.ResultsOur studies indicate that changes in chlorophyll fluorescence lifetime can be utilised as an indirect readout of a photosystem II inhibiting herbicide activity in living plant leaves at three different scales: cellular (~μm), single point (~1 mm2) and macroscopic (~8 × 6 mm2 of a leaf). Multiphoton excited fluorescence lifetime imaging of Triticum aestivum leaves indicated that there is an increase in the spatially averaged chlorophyll fluorescence lifetime of leaves treated with Flagon EC—a photosystem II inhibiting herbicide. The untreated leaf exhibited an average lifetime of 560 ± 30 ps while the leaf imaged 2 h post treatment exhibited an increased lifetime of 2000 ± 440 ps in different fields of view. The results from in vivo wide-field single-photon excited fluorescence lifetime imaging excited at 440 nm indicated an increase in chlorophyll fluorescence lifetime from 521 ps in an untreated leaf to 1000 ps, just 3 min after treating the same leaf with Flagon EC, and to 2150 ps after 27 min. In vivo single point fluorescence lifetime measurements demonstrated a similar increase in chlorophyll fluorescence lifetime. Untreated leaf presented a fluorescence lifetime of 435 ps in the 440 nm excited chlorophyll channel, CH4 (620–710 nm). In the first 5 min after treatment, mean fluorescence lifetime is observed to have increased to 1 ns and then to 1.3 ns after 60 min. For all these in vivo plant autofluorescence lifetime measurements, the plants were not dark-adapted.ConclusionsWe demonstrate that the local impact of a photosystem II herbicide on living plant leaves can be conveniently mapped in space and time via changes in autofluorescence lifetime, which we attribute to changes in chlorophyll fluorescence. Using portable fibre-optic probe instrumentation originally designed for label-free biomedical applications, this capability could be deployed outside the laboratory for monitoring the distribution of herbicides in growing plants.Electronic supplementary materialThe online version of this article (doi:10.1186/s13007-017-0201-7) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

et al. 2017

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“…FLIM has been previously used to investigate the behaviour of intracellular copper(II) [23–27] . In contrast, to the best of our knowledge there are no previous studies where this approach has been employed to visualize copper(I) in live cells.…”

Section: Methodsmentioning

confidence: 99%

“…FLIM has been previously used to investigate the behaviour of intracellular copper(II). [23][24][25][26][27] In contrast, to the best of our knowledge there are no previous studies where this approach has been employed to visualize copper(I) in live cells. Herein we report a new fluorescent probe (FLCS1, Scheme 1), which switches-on its intensity in the presence of copper(I).…”

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confidence: 99%

Selective Detection of Cu⁺ Ions in Live Cells via Fluorescence Lifetime Imaging Microscopy

et al. 2021

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Copper is an essential trace element in living organisms with its levels and localisation being carefully managed by the cellular machinery. However, if misregulated, deficiency or excess of copper ions can lead to several diseases. Therefore, it is important to have reliable methods to detect, monitor and visualise this metal in cells. Herein we report a new optical probe based on BODIPY, which shows a switchon in its fluorescence intensity upon binding to copper(I), but not in the presence of high concentration of other physiologically relevant metal ions. More interestingly, binding to copper(I) leads to significant changes in the fluorescence lifetime of the new probe, which can be used to visualize copper(I) pools in lysosomes of live cells via fluorescence lifetime imaging microscopy (FLIM).

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“…With the FLIM technique, the uptake and release of Cu 2+ in plant cells were visualized through the fluorescence lifetime of GFP. 87,97 Among the genetically encoded lifetime biosensors, most are designed for the evaluation or analysis of redox landscapes and dynamics. The cellular redox states are mainly regulated by pyridine nucleotides (NADH/NAD + and NADPH/NADP + ), thiols and reactive oxygen species, and play vital roles in cell metabolism and cellular signaling.…”

Section: Analytesmentioning

confidence: 99%

“…With the FLIM technique, the uptake and release of Cu 2+ in plant cells were visualized through the fluorescence lifetime of GFP. 87,97…”

Section: Applications Of Fluorescence Lifetime Biosensorsmentioning

confidence: 99%

Genetically encoded fluorescence lifetime biosensors: overview, advances, and opportunities

Mo,

Zhou,

et al. 2023

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Investigation of copper homeostasis in plant cells by fluorescence lifetime imaging microscopy

Cited by 7 publications

References 23 publications

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

Selective Detection of Cu⁺ Ions in Live Cells via Fluorescence Lifetime Imaging Microscopy

Genetically encoded fluorescence lifetime biosensors: overview, advances, and opportunities

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Investigation of copper homeostasis in plant cells by fluorescence lifetime imaging microscopy

Cited by 7 publications

References 23 publications

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

In vivo label-free mapping of the effect of a photosystem II inhibiting herbicide in plants using chlorophyll fluorescence lifetime

Selective Detection of Cu+ Ions in Live Cells via Fluorescence Lifetime Imaging Microscopy

Genetically encoded fluorescence lifetime biosensors: overview, advances, and opportunities

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Selective Detection of Cu⁺ Ions in Live Cells via Fluorescence Lifetime Imaging Microscopy