Confocal laser-scanning microscopy images were obtained using either a Zeiss LSM 880 (with Zen 2.1 SP3 Black edition), Leica SP8 (with LasX 3.1.5.16308) or Leica SP8-MP (with LasX 3.5.6.21594) microscopes. For green and red fluorophores, the following excitation and detection windows were used: mVENUS/GFP/FY/FDA 488 nm, 500-530 nm; mCITRINE 496 nm, 505-530 nm; PI 520 nm, 590-650 nm; Calcofluor White 405nm, 430-485 nm; Basic Fuchsin/Nile Red 561nm, 600-630 nm. For multiphoton microscopy the following excitation and detection settings were used: mVENUS/GFP/FY/Calcofluor White 960 nm, 435-485 nm (Calcofluor White) and 500-550 nm (mVENUS/GFP/FY). Methods for imaging the CS lignin and PI penetration were previously described25 (link),28 (link). For visualization of FDA transport, chambered cover glasses (Thermo Scientific), were used where the roots were covered with a slice of agar and time lapses were made right after the application of FDA.
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Confocal microscopy imaging protocols
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Agar
Basic fuchsin
Calcofluor white
Confocal laser scanning microscopy
Glasses
Lignin
Microscopy
Roots
Corresponding Organization : University of Lausanne
Other organizations : University of Neuchâtel, SIB Swiss Institute of Bioinformatics, University of Zurich
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Variable analysis
independent variables
- Microscopy technique (confocal laser-scanning microscopy, multiphoton microscopy)
- Fluorophore excitation wavelengths (488 nm, 496 nm, 520 nm, 405 nm, 561 nm, 960 nm)
- Fluorophore detection windows (500-530 nm, 505-530 nm, 590-650 nm, 430-485 nm, 600-630 nm, 435-485 nm, 500-550 nm)
- Fluorescence imaging of mVENUS/GFP/FY/FDA, mCITRINE, PI, Calcofluor White, Basic Fuchsin/Nile Red
- Time-lapse imaging of FDA transport
- Microscope models (Zeiss LSM 880, Leica SP8, Leica SP8-MP)
- Microscope software versions (Zen 2.1 SP3 Black edition, LasX 3.1.5.16308, LasX 3.5.6.21594)
- Chambered cover glasses for FDA transport experiments
- CS lignin and PI penetration imaging methods previously described in references 25 and 28
- Not explicitly mentioned
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