Meglumine

(1,2-distearoyl-sn-glycero-3-phosphocholine) (DSPC) 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (ammonium salt) (mPEG₂₀₀₀-DSPE) and cholesterol were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Bortezomib (BTZ) was purchased from LC Laboratories (Woburn, MA, USA). Tiron, meglumine, mannitol, and glucose were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sepharose CL-4B matrix was purchased from GE Life Sciences (Marlborough, MA, USA). NCI-H929, OPM-2, and Hepa1-6 cells were purchased from ATCC (Manassas, VA, USA). RPMI-1640, DMEM, fetal bovine serum, and penicillin-streptomycin solution were purchased from Gibco/ThermoFisher Scientific (Waltham, MA, USA).

Embryos were dissected in FHM medium (95 mM NaCl; 2.5 mM KCl; 0.35 mM KH₂PO₄; 0.2 mM MgSO₄.7H2O; 10 mM Na lactate; 0.2 mM Na pyruvate; 0.2 mM Glucose; 1.0 mM Glutamine; 0.01 mM EDTA; 4.0 mM NaHCO₃; 1.71 mM CaCl₂.2H₂O; 20 mM HEPES; pH adjusted to pH 7.4; after which 1 g/L BSA and 1x Pen/strep (Cat# GIBCO 15140148) were added).
For imaging with the light sheet MuVi SPIM, embryos were fixed in PBS with 4% PFA (ThermoScientific, Cat# 28908) for 10 min at room temperature (short fixation time to reduce background), mounted in 2% LMP agarose (Thermo Scientific, Cat# R0801) in PBS, and imaged immediately.
For imaging with the light sheet UltraMicroscope or OPT, embryos were fixed in PBS with 4% PFA overnight at 4 °C. They were cleared in UBAS-M (25 wt% Meglumine (Sigma-Aldrich, Cat# M9179), 25 wt% Urea (Sigma-Aldrich, Cat# 15604), 20 wt% 1,3-Dimethyl-2-imidazolidinone (Sigma-Aldrich, Cat# 40727), and 0.2 wt% Triton X-100 (Sigma-Aldrich, Cat# 10789704001)) [43 (link)] in 50 ml per embryo overnight at RT and mounted in 4% LMP agarose in UBAS-M before imaging.