Abstract
Abstract Subdiffraction super‐resolution fluorescence microscopy, or nanoscopy, has seen remarkable developments in the last two decades. Yet, for the visualization of plant cells, nanoscopy is still rarely used. In this study, we established RESOLFT nanoscopy on living green plant tissue. Live‐cell RESOLFT nanoscopy requires and utilizes comparatively low light doses and intensities to overcome the diffraction barrier. We generated a transgenic Arabidopsis thaliana plant line expressing the reversibly switchable fluorescent protein rsEGFP2 fused to the mammalian microtubule‐associated protein 4 (MAP4) in order to ubiquitously label the microtubule cytoskeleton. We demonstrate the use of RESOLFT nanoscopy for extended time‐lapse imaging of cortical microtubules in Arabidopsis leaf discs. By combining our approach with fluorescence lifetime gating, we were able to acquire live‐cell RESOLFT images even close to chloroplasts, which exhibit very strong autofluorescence. The data demonstrate the feasibility of subdiffraction resolution imaging in transgenic plant material with minimal requirements for sample preparation.
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Claims 6
Claim · model-written summary
Live-cell RESOLFT nanoscopy was established on living green plant tissue.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianat0 - Outcome
- RESOLFT nanoscopye0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e0“we established RESOLFT nanoscopy on living green plant tissue”
Claim · model-written summary
Live-cell RESOLFT nanoscopy uses comparatively low light doses and intensities to overcome the diffraction barrier.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Outcome
- light doses and intensitiese1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e1“requires and utilizes comparatively low light doses and intensities to overcome the diffraction barrier”
Claim · model-written summary
A transgenic Arabidopsis thaliana plant line expressing the switchable fluorescent protein rsEGFP2 was generated.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Material
- transgenic Arabidopsis thaliana plant linee2
- Gene
- rsEGFP2e2
- Outcome
- expressing the reversibly switchable fluorescent protein rsEGFP2e2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e2“We generated a transgenic Arabidopsis thaliana plant line expressing the reversibly switchable fluorescent protein rsEGFP2”
Claim · model-written summary
Extended imaging of cortical microtubules in Arabidopsis leaf discs was demonstrated with RESOLFT nanoscopy.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Organ / stage
- leaf discse4
- Outcome
- imaging of cortical microtubulese4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e4“imaging of cortical microtubules in Arabidopsis leaf discs”
Claim · model-written summary
Combining RESOLFT nanoscopy with fluorescence lifetime gating allowed live-cell images close to chloroplasts despite their very strong autofluorescence.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Organ / stage
- chloroplastse6
- Treatment
- fluorescence lifetime gatinge5
- Outcome
- RESOLFT imagese6
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e5“By combining our approach with fluorescence lifetime gating”
e6“we were able to acquire live‐cell RESOLFT images even close to chloroplasts, which exhibit very strong autofluorescence”
Claim · model-written summary
Subdiffraction-resolution imaging is feasible in transgenic plant material with minimal sample preparation.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Material
- transgenic plant materiale7
- Outcome
- feasibility of subdiffraction resolution imaginge7
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“transgenic Arabidopsis thaliana”
e7“The data demonstrate the feasibility of subdiffraction resolution imaging in transgenic plant material with minimal requirements for sample preparation”