Imaging Vacuolar Anthocyanins with Fluorescence Lifetime Microscopy (FLIM)
Alexandra Chanoca1,2, Brian Burkel3, Erich Grotewold4
1Department of Botany, University of Wisconsin-Madison, Madison, WI, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 20, 2018
Summary
We developed a fluorescence imaging platform to study plant pigments called anthocyanins. This technique reveals anthocyanin location and vacuolar pH within plant cells.
Area of Science:
- Plant biology
- Biochemistry
- Microscopy
Background:
- Anthocyanins are naturally fluorescent pigments found in plant vacuoles.
- Fluorescence lifetime is sensitive to environmental factors like pH and oxygen concentration.
Purpose of the Study:
- To develop and apply a fluorescence lifetime imaging microscopy (FLIM) platform for analyzing anthocyanins.
- To investigate the subcellular distribution and environmental influences on anthocyanins in plant cells.
Main Methods:
- Utilized FLIM to measure fluorescence decay of anthocyanins in vitro and in vivo.
- Correlated fluorescence lifetime with subcellular localization (vacuolar vs. cytoplasmic) and solution pH.
Main Results:
- Anthocyanin fluorescence lifetime differed between subcellular compartments, with vacuolar anthocyanins showing shorter lifetimes.
- Lower pH in anthocyanin solutions correlated with shorter fluorescence lifetimes.
- FLIM successfully visualized anthocyanin distribution within intact plant cells.
Conclusions:
- FLIM is a valuable tool for studying anthocyanin subcellular distribution.
- Anthocyanin fluorescence lifetime can serve as an indicator of vacuolar pH variations in living cells.
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