Microscopy:Article Title: Quantification of microglial contact and engulfment of oligodendrocyte progenitor cells in the rodent brain.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-Iba-1 (1:750) Wako Cat# 019-19741 Rat anti-PDGFRa (1:400) BD Bioscience Cat# 558774 Rabbit anti-NG2 (1:200 free-floating) Millipore Cat# ab5320 Rabbit anti-Olig2 (1:200) Millipore Cat# ab9610 Alexa Fluor 488 goat anti-rabbit IgG (1:1000) Invitrogen Cat# A11034 Alexa Flour 647 goat anti-rat (1:1000) Invitrogen Cat# A21247 Chemicals, peptides, and recombinant proteins Sodium chloride (NaCl) Sigma-Aldrich Cat #71376 Potassium chloride (KCl) Sigma-Aldrich Cat #P9541 Sodium phosphate dibasic dihydrate (Na2HPO4$2H2O) Sigma-Aldrich Cat #71643 Potassium phosphate monobasic (KH2PO4) Sigma-Aldrich Cat #P9791 Paraformaldehyde (PFA) Sigma-Aldrich Cat #158127 Sucrose Sigma-Aldrich Cat # S0389 O.C.T. .. Compound Fisher Healthcare Cat #23-730-571 Bisbenzimide dye Invitrogen Cat # H3569 Immersion oil Nikon n/a Triton X-100 Sigma-Aldrich Cat #T8787 Experimental models: organisms/strains Mouse: Tg(Cspg4-DsRed.T1)1Akik/J (NG2DsRedBAC), aged 2–8 months, male and female for breeding Jackson Laboratory Cat# Jax 008241, RRID: IMSR_JAX:008241 Mouse: B6.129P2(Cg)-Cx3cr1tm1Litt/J (CX3CR1-GFP), aged 2–8 months, male and female for breeding Jackson Laboratory Cat# Jax 005582, RRID: IMSR_JAX:005582 Software and algorithms Imaris software equipped with the Surface-Surface Coloc XTension Bitplane https://imaris.oxinst.com/ Microsoft Excel Microsoft Corporation https://products.office.com/en/excel MATLAB MathWorks https://www.mathworks.com/ NIS-Elements Nikon https://www.microscope.healthcare. nikon.com/products/software/nis-elements Other Cryostat Leica Biosystems n/a Fluorescent Confocal Microscope with 403 oil immersion objective lens Nikon n/a Fluoromount-G SouthernBiotech Cat #0100-01 Glass coverslips Fisher Scientific Cat #125485MP Sterile Vacuum Filters Fisher Scientific Cat #566-0020 Isoflurane vaporizer system for small animal anesthesia VetEquip http://vetequip.com Brain embedding mold Thermo Scientific Cat #1830 Glass microscopy slides Fisher Scientific Cat #22-037-246 Peristaltic perfusion pump Fisher Scientific Cat # 13-876-1 Sharp scissors Fine Science Tools Cat #14060-09 Forceps Fine Science Tools Cat #11051-10 2–3 Hemostats Fine Science Tools Cat #13008-12 Bone rongeur Fine Science Tools Cat #16000-14 Spatula Fine Science Tools Cat #10099-15 ..
Article Title: Protocol for visualizing microglial lysosomal content by immunohistochemistry in the rodent brain.
Article Snippet: Charles River Cat# T-273; RRID: 2308816 Mouse: C57BL/6J (wild type) males and females aged 4–8 weeks. .. The Jackson Laboratory Cat# 000664; RRID: IMSR_JAX:000664 Software and algorithms IMARIS Software (Version 10.1.1) Oxford Instruments RRID: SCR_007370 Zeiss Zen Black Zeiss RRID: SCR_018163 FIJI NIH RRID: SCR_002285; https://imagej.net/ij/download.html Other Sodium borohydride (NaBH 4 ), granular, 10–40 mesh, 98% Sigma Aldrich Cat#: 452874; CAS: 16940-66-2 Sodium azide BioShop Cat#: SAZ001; CAS: 26628-22-8 Triton X-100 BioShop Cat#: TRX777; CAS: 9002-93-1 MES BioShop Cat#: M3671; CAS: 4432-31-9 NaCl BioShop Cat#: SOD001; CAS: 7647-14-5 EGTA BioShop Cat#: EGT202; CAS: 67-42-5 MgCl 2 ⋅6H 2 O BioShop Cat#: MAG510; CAS: 7791-18-6 Glucose BioShop Cat#: GLU501; CAS: 50-99-7 Normal Goat Serum Millipore Sigma Cat#: S26-M-100ML ProLongTM Gold Antifade Mountant Medium Thermo Fisher Scientific Cat#: P36930 No. 11⁄2 Thickness High Performance Cover Glasses (18 mm x 18 mm) Zeiss Cat#: 474030-9000-000 FisherbrandTM SuperfrostTM Microscope Slides White Tab Fisher Scientific Cat#: 12-550-143 CoverGripTM Coverslip Sealant Biotium Cat#: 23005-1 Corning® LSETM Platform Rocker Millipore Sigma Cat#: CLS6704 Zeiss LSM 880 with Airyscan Confocal Laser Scanning Microscope Zeiss RRID: SCR_020925 Immersion Oil Immersol 518F Zeiss Cat#: 433802-9010-000 Stainless Steel Surgical Prep Blades (for Vibratome) Personna Cat#: 74-0002 Triton X-100 Stock Solution (20% Triton X-100) Reagent Final concentration Amount Triton X-100 20% 10 ml PBS 1X 40 ml Total N/A 50 ml Store at 4 ◦ C long-term (up to 6 months). ..
Sterility:Article Title: Quantification of microglial contact and engulfment of oligodendrocyte progenitor cells in the rodent brain.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-Iba-1 (1:750) Wako Cat# 019-19741 Rat anti-PDGFRa (1:400) BD Bioscience Cat# 558774 Rabbit anti-NG2 (1:200 free-floating) Millipore Cat# ab5320 Rabbit anti-Olig2 (1:200) Millipore Cat# ab9610 Alexa Fluor 488 goat anti-rabbit IgG (1:1000) Invitrogen Cat# A11034 Alexa Flour 647 goat anti-rat (1:1000) Invitrogen Cat# A21247 Chemicals, peptides, and recombinant proteins Sodium chloride (NaCl) Sigma-Aldrich Cat #71376 Potassium chloride (KCl) Sigma-Aldrich Cat #P9541 Sodium phosphate dibasic dihydrate (Na2HPO4$2H2O) Sigma-Aldrich Cat #71643 Potassium phosphate monobasic (KH2PO4) Sigma-Aldrich Cat #P9791 Paraformaldehyde (PFA) Sigma-Aldrich Cat #158127 Sucrose Sigma-Aldrich Cat # S0389 O.C.T. .. Compound Fisher Healthcare Cat #23-730-571 Bisbenzimide dye Invitrogen Cat # H3569 Immersion oil Nikon n/a Triton X-100 Sigma-Aldrich Cat #T8787 Experimental models: organisms/strains Mouse: Tg(Cspg4-DsRed.T1)1Akik/J (NG2DsRedBAC), aged 2–8 months, male and female for breeding Jackson Laboratory Cat# Jax 008241, RRID: IMSR_JAX:008241 Mouse: B6.129P2(Cg)-Cx3cr1tm1Litt/J (CX3CR1-GFP), aged 2–8 months, male and female for breeding Jackson Laboratory Cat# Jax 005582, RRID: IMSR_JAX:005582 Software and algorithms Imaris software equipped with the Surface-Surface Coloc XTension Bitplane https://imaris.oxinst.com/ Microsoft Excel Microsoft Corporation https://products.office.com/en/excel MATLAB MathWorks https://www.mathworks.com/ NIS-Elements Nikon https://www.microscope.healthcare. nikon.com/products/software/nis-elements Other Cryostat Leica Biosystems n/a Fluorescent Confocal Microscope with 403 oil immersion objective lens Nikon n/a Fluoromount-G SouthernBiotech Cat #0100-01 Glass coverslips Fisher Scientific Cat #125485MP Sterile Vacuum Filters Fisher Scientific Cat #566-0020 Isoflurane vaporizer system for small animal anesthesia VetEquip http://vetequip.com Brain embedding mold Thermo Scientific Cat #1830 Glass microscopy slides Fisher Scientific Cat #22-037-246 Peristaltic perfusion pump Fisher Scientific Cat # 13-876-1 Sharp scissors Fine Science Tools Cat #14060-09 Forceps Fine Science Tools Cat #11051-10 2–3 Hemostats Fine Science Tools Cat #13008-12 Bone rongeur Fine Science Tools Cat #16000-14 Spatula Fine Science Tools Cat #10099-15 ..
Laser-Scanning Microscopy:Article Title: Protocol for visualizing microglial lysosomal content by immunohistochemistry in the rodent brain.
Article Snippet: Charles River Cat# T-273; RRID: 2308816 Mouse: C57BL/6J (wild type) males and females aged 4–8 weeks. .. The Jackson Laboratory Cat# 000664; RRID: IMSR_JAX:000664 Software and algorithms IMARIS Software (Version 10.1.1) Oxford Instruments RRID: SCR_007370 Zeiss Zen Black Zeiss RRID: SCR_018163 FIJI NIH RRID: SCR_002285; https://imagej.net/ij/download.html Other Sodium borohydride (NaBH 4 ), granular, 10–40 mesh, 98% Sigma Aldrich Cat#: 452874; CAS: 16940-66-2 Sodium azide BioShop Cat#: SAZ001; CAS: 26628-22-8 Triton X-100 BioShop Cat#: TRX777; CAS: 9002-93-1 MES BioShop Cat#: M3671; CAS: 4432-31-9 NaCl BioShop Cat#: SOD001; CAS: 7647-14-5 EGTA BioShop Cat#: EGT202; CAS: 67-42-5 MgCl 2 ⋅6H 2 O BioShop Cat#: MAG510; CAS: 7791-18-6 Glucose BioShop Cat#: GLU501; CAS: 50-99-7 Normal Goat Serum Millipore Sigma Cat#: S26-M-100ML ProLongTM Gold Antifade Mountant Medium Thermo Fisher Scientific Cat#: P36930 No. 11⁄2 Thickness High Performance Cover Glasses (18 mm x 18 mm) Zeiss Cat#: 474030-9000-000 FisherbrandTM SuperfrostTM Microscope Slides White Tab Fisher Scientific Cat#: 12-550-143 CoverGripTM Coverslip Sealant Biotium Cat#: 23005-1 Corning® LSETM Platform Rocker Millipore Sigma Cat#: CLS6704 Zeiss LSM 880 with Airyscan Confocal Laser Scanning Microscope Zeiss RRID: SCR_020925 Immersion Oil Immersol 518F Zeiss Cat#: 433802-9010-000 Stainless Steel Surgical Prep Blades (for Vibratome) Personna Cat#: 74-0002 Triton X-100 Stock Solution (20% Triton X-100) Reagent Final concentration Amount Triton X-100 20% 10 ml PBS 1X 40 ml Total N/A 50 ml Store at 4 ◦ C long-term (up to 6 months). ..
Concentration Assay:Article Title: Protocol for visualizing microglial lysosomal content by immunohistochemistry in the rodent brain.
Article Snippet: Charles River Cat# T-273; RRID: 2308816 Mouse: C57BL/6J (wild type) males and females aged 4–8 weeks. .. The Jackson Laboratory Cat# 000664; RRID: IMSR_JAX:000664 Software and algorithms IMARIS Software (Version 10.1.1) Oxford Instruments RRID: SCR_007370 Zeiss Zen Black Zeiss RRID: SCR_018163 FIJI NIH RRID: SCR_002285; https://imagej.net/ij/download.html Other Sodium borohydride (NaBH 4 ), granular, 10–40 mesh, 98% Sigma Aldrich Cat#: 452874; CAS: 16940-66-2 Sodium azide BioShop Cat#: SAZ001; CAS: 26628-22-8 Triton X-100 BioShop Cat#: TRX777; CAS: 9002-93-1 MES BioShop Cat#: M3671; CAS: 4432-31-9 NaCl BioShop Cat#: SOD001; CAS: 7647-14-5 EGTA BioShop Cat#: EGT202; CAS: 67-42-5 MgCl 2 ⋅6H 2 O BioShop Cat#: MAG510; CAS: 7791-18-6 Glucose BioShop Cat#: GLU501; CAS: 50-99-7 Normal Goat Serum Millipore Sigma Cat#: S26-M-100ML ProLongTM Gold Antifade Mountant Medium Thermo Fisher Scientific Cat#: P36930 No. 11⁄2 Thickness High Performance Cover Glasses (18 mm x 18 mm) Zeiss Cat#: 474030-9000-000 FisherbrandTM SuperfrostTM Microscope Slides White Tab Fisher Scientific Cat#: 12-550-143 CoverGripTM Coverslip Sealant Biotium Cat#: 23005-1 Corning® LSETM Platform Rocker Millipore Sigma Cat#: CLS6704 Zeiss LSM 880 with Airyscan Confocal Laser Scanning Microscope Zeiss RRID: SCR_020925 Immersion Oil Immersol 518F Zeiss Cat#: 433802-9010-000 Stainless Steel Surgical Prep Blades (for Vibratome) Personna Cat#: 74-0002 Triton X-100 Stock Solution (20% Triton X-100) Reagent Final concentration Amount Triton X-100 20% 10 ml PBS 1X 40 ml Total N/A 50 ml Store at 4 ◦ C long-term (up to 6 months). ..
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