Mouse tissue was fixed with 4% PFA, cryopreserved, and processed with Leica CM3050 S to 8 μm slice. The sections were blocked with 5% donkey serum/0.3% Triton X-100 and incubated with primary antibody against VE-Cadherin (R&D Systems), IGF1R and phosphor-IGF1R (Cell Signaling Technology), and Ki67 (DAKO). Sections were then incubated with fluorescence-labeled secondary antibody (Jackson ImmunoResearch), followed by counterstaining with DAPI (Invitrogen). Images were captured with AxioVert LSM710 confocal microscope (Zeiss).
Axiovert lsm710 confocal microscope
Manufactured by Zeiss
The AxioVert LSM710 is a confocal microscope manufactured by Zeiss. It is a high-resolution imaging system that uses a focused laser beam to scan a sample and capture detailed, three-dimensional images. The microscope is designed to provide high-quality, optical sectioning of specimens.
Automatically generated - may contain errors
Lab products found in correlation
Cm3050, by Leica (1 mentions)
Fluorescence labeled secondary antibody, by Jackson ImmunoResearch (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ve cadherin, by R&D Systems (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Prolong gold antifade solution, by Thermo Fisher Scientific (1 mentions)
Normal donkey serum (nds), by Jackson ImmunoResearch (1 mentions)
Ab90716, by Abcam (1 mentions)
Click it plus edu imaging kit, by Thermo Fisher Scientific (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
3 protocols using axiovert lsm710 confocal microscope
1
Multimodal Immunofluorescence of Mouse Tissue
2
Quantifying Lung Cell Proliferation
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Lung cell proliferation was measured in vivo by EdU labeling. Briefly, 100 µg of EdU in 200 µl PBS was intraperitoneally injected once a day from day 7 to day 9. At day 10, lungs were fixed with 4% PFA and embedded in optimum cutting temperature formulations (Tissue-Tek). 14-µm-thick cryosections were incubated with PBS containing 5% normal donkey serum (Jackson ImmunoResearch) and 0.15% Triton X-100 (Sigma-Aldrich) for blocking and permeation. EdU was developed with Click-iT Plus EdU Imaging Kits (Molecular Probes; C10640), and sequentially, sections were incubated with diluted primary antibodies in a humidified chamber overnight at 4°C. Primary antibodies against Prosurfactant Protein C (Abcam; ab90716, 1:800), Desmin (Abcam; ab15200, 1:200), and VE-cadherin (Biolegend; BV13, 1:100) were used. 24 h later, the cryosections were incubated with secondary antibody (Thermo Fisher Scientific) for 1 h under ambient temperature. The slides were mounted using Prolong Gold antifade solution (Invitrogen). Images were captured on an AxioVert LSM710 confocal microscope (Zeiss).
3
Histological Analysis of Lung Slides
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