Journal: Journal of visualized experiments : JoVE

Article Title: Analysis of Non-Human Primate Pancreatic Islet Oxygen Consumption

doi: 10.3791/60696

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Article Snippet: We utilized a 0.45 μm filter ( ). table ft1 table-wrap mode="anchored" t5 Name Company Catalog Number Comments Cell culture dish, 60 mm X 15 mm style Corning 430166 Cell-Tak Cell and Tissue Adhesive Corning 354240 Conical tube, 50 mL Falcon 352070 Dextrose anhydrous Fisher Scientific BP350–1 For glucose solution, 200 mg/ml, sterile filetered Disposable reservoirs (sterile), 25 ML Vistalab 3054–1033 for loading multichannel pipet EZFlow Sterile 0.45 μm PES Syringe Filter, 13 mm Foxx Life Sciences 371–3115-OEM L-glutamine Gibco 25030–081 200 mM (100x) Multichannel pipette tips ThermoFisher Scientific 94410810 Multichannel pipette, 15–1250 μL ThermoFisher Scientific 4672100BT Recommended P20, P200, and P1000 pipettes Eppendorf 2231000602 pH Probe Hanna Instruments HI2210–01 Pipette tips, 20 μL, 200 μL, 1000 μL Olympus 24–404, 24–412, 24–430 Seahorse XF Base Media Agilent 103334–100 Seahorse XF Cell Mito Stress Test Kit Agilent 103015–100 Includes Oligomycin, FCCP, and Rotenone/Antimycin A Seahorse XFe96 Analyzer Agilent S7800B Including prep station with 37 °C non-CO2 incubator Seahorse XFe96 Spheroid Fluxpak Mini Agilent 102905–100 Includes sensor cartridge, spheroid microplate, and calibrant Sodium bicarbonate Fisher Scientific BP328–500 Sodium pyruvate Gibco 11360–070 100 mM (100x) Stereo Microscope Olympus SZX9 Syringe (sterile), 5 mL BD 309603 For sterile filtration Water (sterile) Sigma W3500–500mL Open in a separate window In a cell culture hood, add 200 μL of cell and tissue adhesive ( ) to 2.8 mL of 0.1M sodium bicarbonate.

Techniques: Cell Culture, Adhesive, Sterility, Transferring, Microscopy, Filtration