Isolation of Single Intracellular Bacterial Communities Generated from a Murine Model of Urinary Tract Infection for Downstream Single-Cell Analysis

Journal of Visualized Experiments www.jove.com

Materials List for: Isolation of Single Intracellular Bacterial Communities Generated from a Murine Model of Urinary Tract Infection for Downstream Single-cell Analysis

EnJun Yang1, Jacqueline L.Y. Chee1, Suhanya Duraiswamy2, Siyi Chen3, Kristin Lees3, Swaine L. Chen1,3 1 Infectious Diseases Group, Genome Institute of Singapore 2 Department of Chemical and Biomolecular Engineering, National University of Singapore 3 Division of Infectious Diseases, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore

Correspondence to: Swaine L. Chen at [email protected]

URL: https://www.jove.com/video/58829 DOI: doi:10.3791/58829

Materials

Name Company Catalog Number Comments 1.5 mL eppendorf tube For static bacterial culture and OD measurement 100% ethanol For Alcohol Burner 15 mL conical tube For static bacterial culture and OD measurement 1 mL Tuberculin Syringe BD Biosciences 302100 3% Bacterial Agar For static bacterial culture and OD measurement 70% ethanol For static bacterial culture and OD measurement Aesculap anatomic forceps Braun/Kruuse BD222R For initial dissection of mouse (skin, fascia) Alcohol Burner Wheaton 237070 Aspirating pipette BD Biosciences 357558 Aspirator tube Sigma-Aldrich A5177 Bacterial loops For static bacterial culture and OD measurement Benchtop centrifuge Eppendorf 5424 Any centrifuge for 1.5ml eppendorf tubes Conical flasks For static bacterial culture and OD measurement Digital camera for microscope Olympus DP71 For image capture and harvesting of IBCs. Any other fluorescent microscope with a GFP channel will suffice Glass Capillaries Kimax 6148K07 Iris Scissors STR SS 110MM Braun BC110R Isoflurane (Isothesia) Henry Schein Animal Health 29405 Kanamycin Sulfate Calbiochem 420311 For static bacterial culture and OD measurement LB broth (Miller) Thermo/Gibco 10855021 For static bacterial culture and OD measurement Light source unit for microscope Olympus LG-PS2 For image capture and harvesting of IBCs. Any other fluorescent microscope with a GFP channel will suffice Lubricant KY Any similar commercial medical lubricant will suffice

Macro fluorescence microscope Olympus MVX10 For image capture and harvesting of IBCs. Any other fluorescent microscope with a GFP channel will suffice Micropipette + micropipette tips For static bacterial culture and OD measurement PBS 1x For static bacterial culture and OD measurement Pipette controller + Pipettes For static bacterial culture and OD measurement Polyethylene Tubing BD Intramedic 427401 Precision Glide needle 30 G BD Biosciences 305107 Possibly under new catalogue number (305106) Splinter forceps curved Braun BD312R Spray bottle (for ethanol) For static bacterial culture and OD measurement Square cuvettes Elkay 127-1010-400 For static bacterial culture and OD measurement Sterilgard III Advance Safety Baker SG403 Any biosafety cabinet with a UV Cabinet irridiator Sterilin 90mm Standard Petri Dish Thermo 101VR20 Any sterile petri dish Stevens, vascular and tendon Braun OK366R Recommended for harvesting of scissors, curved, delicate, 110 mm bladder Surgical Scissors STR S/B 105MM Braun BC320R Tabletop Centrifuge Eppendorf 5810R Any refridgerated centrifuge for 15ml conicals WPA C08000 cell density meter Biowave (Biochrom) 80-3000-45 For static bacterial culture and OD measurement