Adult male and female RjOrl:SWISS mice (2 months old) were transcardially perfused with 0.9% saline and 3.2% paraformaldehyde. Brains were fixed overnight and 20 μm coronal sections cut with a Leica VT1000S vibrating microtome. Brain sections were incubated in blocking solution (PBS with 2% bovine serum albumin (BSA) and 0.2% (Ki67 and DCX) or 0.05% (NG2) Triton X-100) for 60 minutes and then incubated in primary antibodies in blocking solution for 36 hours at 4°C. After washing, sections were revealed with Alexa Fluor-conjugated secondary antibodies (Life Technologies and Jackson ImmunoResearch Laboratories) for 1–2 hours at room temperature, counterstained with 4’,6-Diamidino-2-Phenylindole Dihydrochloride (DAPI), (1 μg/ml, 5 minutes, SIGMA D9542) and mounted with FluorSave™ (Millipore corporation). The following primary antibodies were used: anti-rabbit NG2 (Merck AB5320, 1:100), anti-rabbit Iba1 (Wako Cat. #019–19741, 1:100), anti-rabbit Ki67 (Abcam ab15580, 1:100), anti-goat doublecortin (Santa Cruz (C18): sc-8066, 1:150). Samples were imaged using an inverted LSM 700 confocal (Zeiss).
Inverted lsm 700 confocal
Manufactured by Zeiss
The Inverted LSM 700 confocal is a high-performance laser scanning microscope designed for advanced imaging applications. It features a compact and versatile inverted configuration, allowing for easy sample handling and observation. The system utilizes a powerful laser source and state-of-the-art detector technology to provide high-resolution, high-quality images with excellent signal-to-noise ratio. The Inverted LSM 700 confocal is capable of performing a wide range of imaging techniques, including fluorescence, reflectance, and transmitted light microscopy.
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2 protocols using inverted lsm 700 confocal
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Immunolabeling of Neural Stem Cells
2
Immunolabeling of Neural Stem Cells
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Adult male and female RjOrl:SWISS mice (2 months old) were transcardially perfused with 0.9% saline and 3.2% paraformaldehyde. Brains were fixed overnight and 20 μm coronal sections cut with a Leica VT1000S vibrating microtome. Brain sections were incubated in blocking solution (PBS with 2% bovine serum albumin (BSA) and 0.2% (Ki67 and DCX) or 0.05% (NG2) Triton X-100) for 60 minutes and then incubated in primary antibodies in blocking solution for 36 hours at 4°C. After washing, sections were revealed with Alexa Fluor-conjugated secondary antibodies (Life Technologies and Jackson ImmunoResearch Laboratories) for 1–2 hours at room temperature, counterstained with 4’,6-Diamidino-2-Phenylindole Dihydrochloride (DAPI), (1 μg/ml, 5 minutes, SIGMA D9542) and mounted with FluorSave™ (Millipore corporation). The following primary antibodies were used: anti-rabbit NG2 (Merck AB5320, 1:100), anti-rabbit Iba1 (Wako Cat. #019–19741, 1:100), anti-rabbit Ki67 (Abcam ab15580, 1:100), anti-goat doublecortin (Santa Cruz (C18): sc-8066, 1:150). Samples were imaged using an inverted LSM 700 confocal (Zeiss).
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