To monitor the proliferation of the lymphatic vessels 50 mg/kg EdU was injected intraperitoneally 4 days after 1 µg of Poly(C) RNA-LNP or VEGFC mRNA-LNP treatment of the ear of mice. The samples were harvested 24 h after EdU treatment, and histological procedures were performed. The slides were stained with Click-iT EdU Cell Proliferation Kit for Imaging, Alexa Fluor 594 dye (Invitrogen, C10337), anti-LYVE1 antibody (R&D systems, AF2125), and anti-α-SMA antibody (Abcam, ab124964). Confocal images were acquired using a Nikon Eclipse A1 Confocal Laser Microscope (Nikon Instruments) using 10x or 40x dry objectives.
Eclipse a1 confocal laser microscope
Manufactured by Nikon
The Nikon Eclipse A1 Confocal Laser Microscope is a high-performance imaging system designed for advanced microscopy applications. It utilizes a laser-scanning system to provide high-resolution, three-dimensional imaging capabilities. The instrument is equipped with multiple laser lines and a sensitive detector to capture detailed images of samples.
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Lab products found in correlation
Click it edu cell proliferation kit for imaging, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 dye, by Thermo Fisher Scientific (1 mentions)
Anti lyve 1 antibody, by R&D Systems (1 mentions)
Anti α sma antibody, by Abcam (1 mentions)
Ds ri2 camera, by Nikon (1 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
2 protocols using eclipse a1 confocal laser microscope
1
Lymphatic Vessel Proliferation Assay
2
Lymphatic Vessel Visualization in Mice
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The 2% paraformaldehyde (Sigma-Aldrich) fixed ears of Prox1GFP lymphatic reporter mice injected with Poly(C) RNA-LNP or VEGFC mRNA-LNPs were imaged for GFP fluorescence and anti-von Willebrand Factor (anti-vWF) (Sigma-Aldrich, F3520, 1:100) labeling after overnight incubation, which was followed by anti-rabbit secondary antibody staining conjugated to Alexa Fluor 568 (Life Technologies, AF11036, 1:250). Thereafter confocal microscopy was performed using a Nikon Eclipse A1 Confocal Laser Microscope and a Nikon SMZ25 stereo microscope connected to a Nikon DS-Ri2 camera (Nikon Instruments).
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