Mouse brains were fixed with 4% paraformaldehyde, sectioned by vibratome (Leica) or cryostat (Leica) and stained following established protocols6 (link). For brain slice assays6 (link), 250 μm thick slices of adult mouse brain were prepared with a vibratome (Leica) and placed on top of 0.8 μm pore membranes (Millipore) in brain slice culture medium (DMEM, complete HBSS, 5% FBS, 1mM L-glutamine, 100 IU/mL penicillin, 100 μg/mL streptomycin). 3 × 105 cancer cells were placed on the surface of the slice. After 48 h of incubation, brain slices were fixed with 4% paraformaldehyde, and stained. For immunostaining in chamber slide cultures, cells were fixed with 4% paraformaldehyde and stained. Antibodies used for immunochemical staining are listed in Supplementary Information . Images were acquired with Zeiss Axio Imager Z1 microscope or Leica SP5 upright confocal microscope, and analyzed with ImageJ, Imaris and Metamorph softwares. Antibodies used for immunostaining are listed in Supplementary Information .
0.8 μm pore membranes
Manufactured by Merck Group
0.8 μm pore membranes are a type of laboratory filtration equipment designed to filter particles and molecules from liquid or gas samples. These membranes have a pore size of 0.8 micrometers, which allows them to effectively separate and retain specific components based on their size.
Automatically generated - may contain errors
2 protocols using 0.8 μm pore membranes
1
Mouse Brain Slice Assay for Cancer Cell Invasion
2
Mouse Brain Slice Assay for Cancer Cell Invasion
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Mouse brains were fixed with 4% paraformaldehyde, sectioned by vibratome (Leica) or cryostat (Leica) and stained following established protocols6 (link). For brain slice assays6 (link), 250 μm thick slices of adult mouse brain were prepared with a vibratome (Leica) and placed on top of 0.8 μm pore membranes (Millipore) in brain slice culture medium (DMEM, complete HBSS, 5% FBS, 1mM L-glutamine, 100 IU/mL penicillin, 100 μg/mL streptomycin). 3 × 105 cancer cells were placed on the surface of the slice. After 48 h of incubation, brain slices were fixed with 4% paraformaldehyde, and stained. For immunostaining in chamber slide cultures, cells were fixed with 4% paraformaldehyde and stained. Antibodies used for immunochemical staining are listed in Supplementary Information . Images were acquired with Zeiss Axio Imager Z1 microscope or Leica SP5 upright confocal microscope, and analyzed with ImageJ, Imaris and Metamorph softwares. Antibodies used for immunostaining are listed in Supplementary Information .
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