Mouse aortas were prepared and embedded in Tissue-Tek (Sakura Finetek), and serial 10-μm-thick frozen tissue sections were prepared. Immunostainings were performed using following marker antibodies: anti-mouse CD3e (145–2C11, BD; 1:100 dilution), anti-mouse Foxp3 (ab75763, Abcam; 1:100 dilution) and DAPI for DNA. Secondary antibodies were used as previously described7 (link). For negative controls, stainings were performed without primary antibodies or isotype controls. Stained sections were analyzed using a Leica SP8 ×3 microscope (Leica microsystems, Germany). Images were acquired with identical microscope settings using sequential channel acquisitions to avoid cross-talk between fluorophores. All images were prepared as TIF files by Fiji (ImageJ, National Institutes of Health) or Leica LAS-X (V3.5) software and exported into Adobe Illustrator CS6 for figure arrangements.
Sp8 3 microscope
Manufactured by Leica
Sourced in Germany
The Leica SP8 ×3 microscope is a high-performance confocal laser scanning microscope. It features advanced optical components and a modular design to provide researchers with a versatile imaging platform for a wide range of applications.
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2 protocols using sp8 3 microscope
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Immunostaining of Aortic Tissue Sections
2
Immunostaining of Aortic Tissue Sections
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Mouse aortas were prepared and embedded in Tissue-Tek (Sakura Finetek), and serial 10-μm-thick frozen tissue sections were prepared. Immunostainings were performed using following marker antibodies: anti-mouse CD3e (145–2C11, BD; 1:100 dilution), anti-mouse Foxp3 (ab75763, Abcam; 1:100 dilution) and DAPI for DNA. Secondary antibodies were used as previously described7 (link). For negative controls, stainings were performed without primary antibodies or isotype controls. Stained sections were analyzed using a Leica SP8 ×3 microscope (Leica microsystems, Germany). Images were acquired with identical microscope settings using sequential channel acquisitions to avoid cross-talk between fluorophores. All images were prepared as TIF files by Fiji (ImageJ, National Institutes of Health) or Leica LAS-X (V3.5) software and exported into Adobe Illustrator CS6 for figure arrangements.
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