The open reading frames (ORFs) of the MdNup54, MdHSP70, and MdKNAT4/6 genes were inserted independently into the pCAMBIA2300-EGFP vector to generate the 35S:MdNup54-EGFP, 35S:MdHSP70-EGFP, and 35S:MdKNAT4/6-EGFP recombinant plasmids, respectively. These recombinant plasmids were inserted independently into Agrobacterium tumefaciens strain GV3101 cells. Then they were infiltrated into tobacco leaves. GV3101 cells containing the pCAMBIA2300-EGFP vector (35S:EGFP) served as the control. After an additional 3 days of growth in the dark, green fluorescent protein (GFP) signals in transformed tobacco leaves were detected using a Leica TCS SP8 SR Laser Scanning Confocal Microscope (Leica, Germany). The primers used are shown in Supplementary Table 2 .
Tcs sp8 sr laser scanning confocal microscope
Manufactured by Leica
Sourced in Germany
The Leica TCS SP8 SR is a laser scanning confocal microscope designed for high-resolution imaging. It utilizes a scanning laser beam to generate detailed images of samples by collecting light from a specific focal plane, allowing for the reduction of out-of-focus information. The instrument is capable of providing optical sectioning and three-dimensional reconstruction of samples.
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Lab products found in correlation
4 protocols using tcs sp8 sr laser scanning confocal microscope
1
Investigating Gene Function in Tobacco Leaves
2
Transient Expression of Plant Transcription Factors
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The open reading frames (ORFs) of the MdNup62, MdHSFA1d, and MdHSFA9b genes were inserted independently into the pCAMBIA2300-EGFP vector to generate the 35S::MdNup62-EGFP, 35S::MdHSFA1d-EGFP, and 35S::MdHSFA9b-EGFP recombinant plasmids, respectively. These recombinant plasmids were inserted independently into Agrobacterium tumefaciens strain GV3101 cells. The GV3101 cells containing these recombinant plasmids were then infiltrated into tobacco leaves. GV3101 cells containing the pCAMBIA2300-EGFP vector (35S::EGFP) served as the control. After an additional 3 days of growth in the dark, green fluorescent protein (GFP) signals in transformed tobacco leaves were detected using a Leica TCS SP8 SR Laser Scanning Confocal Microscope (Leica, Germany).The primers used are listed in Table S5.
3
Visualizing Subcellular Localization of Fusion Proteins in Tobacco Leaves
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The ORFs of GausACLB-2 amplified from G. australe were fused in GFP into a binary vector pBIN-GFP4. AtPIP2A fusion RFP protein was used as plasma membrane marker [71 (link)]. The excitation wavelengths for imaging GFP and RFP fusions were 488 and 580 nm, respectively. The recombinant vector was then transformed into tobacco (N. benthamiana) leaf epidermal cells as described by Lu et al. [72 (link)]. Fluorescent signals were recorded and visualized by using a Leica TCS SP8 SR Laser scanning confocal microscope (Leica, Germany) with a 488 nm or 580 nm laser, and photographed with a LEICA DFC420 camera under 20X objective lenses. Images were acquired at 1024 × 1024 resolution using LAS X software (Leica Application Suite X) and processed using Adobe Illustrator.
4
Visualizing Transcription Factor Localization
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The open reading frames (ORFs) of the MdNup62 , MdHSFA1d , and MdHSFA9b genes were inserted independently into the pCAMBIA2300-EGFP vector to generate the 35S::MdNup62 -EGFP, 35S::MdHSFA1d -EGFP, and 35S::MdHSFA9b -EGFP recombinant plasmids, respectively. These recombinant plasmids were inserted independently into Agrobacterium tumefaciens strain GV3101 cells. The GV3101 cells containing these recombinant plasmids were then infiltrated into tobacco leaves. GV3101 cells containing the pCAMBIA2300-EGFP vector (35S::EGFP) served as the control. A er an additional 3 days of growth in the dark, green fluorescent protein (GFP) signals in transformed tobacco leaves were detected using a Leica TCS SP8 SR Laser Scanning Confocal Microscope (Leica, Germany). The primers used are listed in Table S5.
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