For light microscopy, a Nikon Eclipse (Nikon, Duesseldorf, Germany) was used. Confocal microscopy images were acquired using a Zeiss LSM710 laser-scanning confocal system with spectral detection capabilities, and Zen 2010 software version 6.0 (Carl Zeiss Microscopy GmbH, Jena, Germany) was used for image acquisition and analysis. Excitation light was provided by 488, 543, and 634 nm laser lines; fluorescence emission was detected at 500–530 nm for Cy2 (green) and 550–600 nm for Cy3 (red) in separate tracks, using a confocal aperture of 1 Airy unit. Some of the images were acquired as z-stacks and 3D reconstruction was performed.
Zen 2010 software version 6
Manufactured by Zeiss
Sourced in Germany
The Zen 2010 software version 6.0 is a core imaging and analysis platform developed by Zeiss. It provides a comprehensive suite of tools for managing, processing, and visualizing image data from various microscopy techniques.
Automatically generated - may contain errors
9 protocols using zen 2010 software version 6
1
Multicolor Confocal Microscopy Imaging
2
Multicolor Confocal Imaging Protocol
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For light microscopy, a Nikon Eclipse (Nikon, Duesseldorf, Germany) was used. Confocal microscopy images were acquired using a Zeiss LSM710 laser-scanning confocal system with spectral detection capabilities, and Zen 2010 software version 6.0 (Carl Zeiss Microscopy GmbH, Jena, Germany) was used for image acquisition and analysis. Excitation light was provided by 488, 543, and 634 nm laser lines; fluorescence emission was detected at 500–530 nm for FITC (green), 550–600 nm for rhodamin (red) and 650–710 nm for Cy5 (blue) in separate tracks, using a confocal aperture of 1 Airy unit. Some of the images were acquired as z-stacks and 3D reconstruction was performed.
3
Multicolor Confocal Microscopy Imaging
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For light microscopy, a Nikon Eclipse (Nikon, Duesseldorf, Germany) was used. Confocal microscopy images were acquired using a Zeiss LSM710 laser-scanning confocal system with spectral detection capabilities, and Zen 2010 software version 6.0 (Carl Zeiss Microscopy GmbH, Jena, Germany) was used for image acquisition and analysis. Excitation light was provided by 488, 543, and 634 nm laser lines; fluorescence emission was detected at 500–530 nm for FITC (green), 550–600 nm for Rhodamine (red), and 650–710 nm for Cy5 (blue) in separate tracks, using a confocal aperture of 1 Airy unit. Some of the images were acquired as z-stacks and 3D reconstruction was performed using a software algorithm (maximal projection).
4
Multicolor Confocal Microscopy Imaging
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Confocal microscopy images were acquired using a Zeiss LSM710 laser-scanning confocal system with spectral detection capabilities, and Zen 2010 software version 6.0 (Carl Zeiss Microscopy GmbH, Jena, Germany) was used for image acquisition and analysis. Excitation light was provided by 488, 543, and 634 nm laser lines; fluorescence emission was detected at 500-530 nm for FITC (green), 550-600 nm for CFL 555 (red) and 360-486 nm for Hoechst (blue) in separate tracks, using a confocal aperture of 1 Airy unit. Some of the images were acquired as z-stacks and 3D reconstruction was performed.
5
Adipocyte Perilipin Expression Dynamics
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Perilipin protein expression was assessed at 3, 7, 14, 21, and 28 days after adipogenic induction using a Carl Zeiss LSM710 confocal microscope. Briefly, both hADSCs/G-A and hADSCs/G-A-PAA biohybrids were fixed with 4% PFA, permeabilized with 2% BSA/0.1% Triton X-100 solution, incubated overnight at 4°C with rabbit polyclonal anti-perilipin antibody solution, and finally exposed to TRITC conjugated goat anti-rabbit secondary antibody. Cell nuclei were stained using DAPI dye for 30 min and the resulting labeled constructs were visualized in confocal fluorescence microscopy using a confocal aperture that corresponded to a back-projected size of 1 airy unit. The 405 and 543 nm laser lines were used for excitation and fluorescence emission was detected at 490–515 nm for DAPI and 600–680 nm for TRITC. Carl Zeiss Zen 2010 software version 6.0 was used for image acquisition and analysis.
6
Visualizing hASC Cytoskeleton on CHT/GO Scaffolds
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F-actin filaments developed by hASCs in contact with each material were studied by confocal microscopy using a Carl Zeiss LSM710 laser-scanning microscope. hASC morphology and cytoskeleton fibers distribution in contact with CHT/GO scaffolds were studied at 48 h post seeding. In order to fluorescently label F-actin, constructs were fixed with 4% paraformaldehyde (PFA) for 8 h and permeabilized with 2% BSA/0.1% Triton X-100 solution at 4 °C. Next, the constructs were incubated 4 h at 37 °C with Phalloidin-FITC (Sigma-Aldrich, Darmstadt, Germany). Cell nuclei were stained with DAPI for 30 min. Carl Zeiss Zen 2010 software version 6.0 was used for image acquisition and analysis.
7
Multicolor Confocal Microscopy Protocol
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For light microscopy, an Axioscope A1 (Zeiss, Germany, Germany) was used. Confocal microscopy images were acquired using a Zeiss LSM710 laser-scanning confocal system with spectral detection capabilities, and Zen 2010 software version 6.0 (Carl Zeiss Microscopy GmbH, Jena, Germany) was used for image acquisition and analysis. Excitation light was provided by 488, 543, and 634 nm laser lines; fluorescence emission was detected at 500–530 nm for FITC (green), 550–600 nm for Rhodamin (red), and 650–710 nm for Cy5 (blue) in separate tracks, using a confocal aperture of 1 Airy unit. Some of the images were acquired as z-stacks and 3D reconstruction was performed using a software algorithm (maximal projection).
8
In Vivo Nanoparticle-Mediated mRNA/sgRNA Delivery
9
In Vivo Nanoparticle-Mediated mRNA/sgRNA Delivery
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Nanoparticles containing Cas9 mRNA and modified sgTom1 (mRNA/sgRNA weight ratio 4:1, total RNA dose 0.75 mg kg−1) were prepared as in vivo iPLNP formulation method mentioned above (Supplementary Table 2 ). 9A1P9: 5A2-SC8: chol: DMG-PEG2000 (molar ratio) of 25:30:30:1 and 9A1P9: DDAB: chol: DMG-PEG2000 (molar ratio) of 60:30:40:0.4 were used for 9A1P9–5A2-SC8 iPLNP and 9A1P9-DDAB iPLNP, respectively. 9A1P9/RNA weight ratio was fixed at 18:1. Afterward, iPLNPs were administered to Ai9 mice via IV injection. PBS group was used as negative control. 10 days later, mice were sacrificed and the organs were isolated and imaged on the IVIS Spectrum in vivo imaging system (Perkin Elmer). Then, tissues were embedded in optimal cutting temperature (OCT) compound and cut into 10 μm sections. After fixing with 4% paraformaldehyde for 20 min, the sections were washed three times with PBS. Afterward, ProLong Gold Mountant with DAPI was dropped to the slides, and coverslips were covered. These slides were imaged by confocal microscopy (Zeiss LSM 700), and analyzed with ZEN 2010 software version 6.0.62 (Carl Zeiss MicroImaging GmbH).
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