Cells were fixed with 4% paraformaldehyde for 20 minutes, permeabilized with 0.2% Triton X-100 in phosphate-buffered saline (PBS) for 30 minutes, and blocked with serum in PBS for 1 h. Cells were then stained with appropriate primary antibodies and AlexaFluor-conjugated secondary antibodies (Life Technologies) and co-stained with DAPI (Vector Laboratories, Burlingame, CA, USA). The primary antibodies for CD31 and CD144 (Abcam, Cambridge, MA, USA) were used in the staining. All murine iECs were also incubated with 10 ug/mL DiI AcLDL (Life Technologies) at 37°C for 6 hours. After washing twice with PBS, DiI AcLDL uptake was detected with fluorescence microscopy. Imaging was performed on a Leica DFC500 inverted microscope.
Dfc500 inverted microscope
Manufactured by Leica
The DFC500 is an inverted microscope designed for high-quality imaging and documentation of biological samples. It features a digital camera for capturing images and a versatile optical system that supports a range of magnification levels and contrast techniques. The core function of the DFC500 is to provide users with a reliable and efficient tool for observing and analyzing their samples.
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2 protocols using dfc500 inverted microscope
1
Immunofluorescence Staining of Endothelial Cells
2
Immunofluorescence Staining of Endothelial Cells
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Cells were fixed with 4% paraformaldehyde for 20 minutes, permeabilized with 0.2% Triton X-100 in phosphate-buffered saline (PBS) for 30 minutes, and blocked with serum in PBS for 1 h. Cells were then stained with appropriate primary antibodies and AlexaFluor-conjugated secondary antibodies (Life Technologies) and co-stained with DAPI (Vector Laboratories, Burlingame, CA, USA). The primary antibodies for CD31 and CD144 (Abcam, Cambridge, MA, USA) were used in the staining. All murine iECs were also incubated with 10 ug/mL DiI AcLDL (Life Technologies) at 37°C for 6 hours. After washing twice with PBS, DiI AcLDL uptake was detected with fluorescence microscopy. Imaging was performed on a Leica DFC500 inverted microscope.
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