The entry vector containing the cds of OsHOL1 was recombined with the p2FGW7 destination vector60 (link) using the Gateway recombination protocol, as previously described, to produce a 35S:GFP-OsHOL1 expression plasmid. Arabidopsis mesophyll protoplasts were isolated and transfected with 5 μg of plasmid, as described in61 (link). An mCherry-NLS–RFP-containing vector62 (link) was used to visualize cell nuclei. For GFP and RFP imaging, Arabidopsis protoplasts were analyzed using a Leica THUNDER Imager Model Organism fluorescence microscope (Leica Microsystems, Germany).
Thunder imager model organism fluorescence microscope
Manufactured by Leica
Sourced in Germany
The Leica THUNDER Imager Model Organism fluorescence microscope is a high-performance imaging system designed for live-cell and fixed-sample imaging. It features advanced optics and a sensitive camera to capture detailed fluorescence images of model organisms.
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2 protocols using thunder imager model organism fluorescence microscope
1
GFP-OsHOL1 Expression in Arabidopsis Protoplasts
2
Visualizing OsHOL1 Protein Interactions
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Vectors for the expression of OsHOL1 proteins fused to the N-terminal or C-terminal YFP fragments were generated by Gateway recombination of the OsHOL1 cds entry vector with the destination vectors pE-SPYNE-GW and pE-SPYCE-GW64 (link),65 (link). Arabidopsis protoplasts, isolated as above described, were transformed with 5 μg of each BiFC vector. An mCherry-NLS–RFP-containing vector62 (link) was used to visualize the cell nuclei. YFP and RFP fluorescence was visualized using a Leica THUNDER Imager Model Organism fluorescence microscope (Leica Microsystems).
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