Four days after agro-infiltration, leaf discs were collected from the plants. Confocal microscopy was performed at the Molecular and Cellular Imaging Center (MCIC), the Ohio Agricultural Research and Development Center, using a Leica DMI6000 laser confocal scanning microscope. To detect GFP and mCherry fluorescence, sequential excitation at 488 nm and 587 nm was provided by argon and helium-neon 543 lasers, respectively.
Dmi6000 laser confocal scanning microscope
Manufactured by Leica
The DMI6000 is a laser confocal scanning microscope manufactured by Leica. It is designed for high-resolution imaging of a variety of samples. The instrument utilizes laser illumination and a pinhole aperture to achieve optical sectioning, enabling the capture of detailed three-dimensional images.
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4 protocols using dmi6000 laser confocal scanning microscope
1
Fluorescence Imaging of Transgenic Plants
2
Fluorescence Imaging of Transgenic Plants
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Four days after agro-infiltration, leaf discs were collected from the plants. Confocal microscopy was performed at the Molecular and Cellular Imaging Center (MCIC), the Ohio Agricultural Research and Development Center, using a Leica DMI6000 laser confocal scanning microscope. To detect GFP and mCherry fluorescence, sequential excitation at 488 nm and 587 nm was provided by argon and helium-neon 543 lasers, respectively.
3
Time-lapse Imaging of Gata6 iPSCs
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Gata6 engineered iPS cells were seeded at 26 K cells per cm2 in a BD six-well tissue culture treated plate coated with BD Matrigel, then induced with 1,000 ng ml−1 Dox. After induction, the cells were viewed under a Leica DMI6000 Confocal Laser Scanning Microscope, in an incubation chamber. Images were taken every 30 min for 4 days.
4
Multicolor Imaging of Intestinal Microbiome
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10 μm thick intestinal tissues sections were deparaffinized in xylol and rehydrated using a descending alcohol series. Then, 500 nM EUB338‐I‐III‐6‐FAM, a 16S rRNA‐targeted DNA oligonucleotide probe mix (Biomers, Ulm, Germany) to detect all bacteria by FISH (for sequences see Table S3 ), were applied in hybridization buffer (900 mM NaCl, 20 mM TRIS‐HCl, pH 7.2, 0.01% SDS) at 46°C for 2 hours and washed in wash buffer (900 mM NaCl, 20 mM TRIS‐HCl, pH 7.2) at 48°C for 10 minutes. For immunofluorescence staining, MUC2 antibody was applied overnight followed by incubation with AlexaFluor 594 secondary antibody (Table S2 ) in wash buffer for 1 hour. To visualize DNA, Hoechst 33258 staining was performed. Slides were embedded in CitiFluor AF1 (Science Services, Munich, Germany). Images were acquired on an inverted DMI 6000 confocal laser scanning microscope (Leica) equipped with a 405 nm UV diode and a Leica supercontinuum white light laser using a ×60 glycerol objective and the software Leica Application Suite AF.
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