Fast dil oil

zCDX models were developed by a CRO (BioReperia AB). Transgenic Tg(fli1:EGFP)y1 zebrafish embryos were raised at 28 °C for 48 h in E3 embryo medium (containing per liter: 0.286 g NaCl, 0.048 g CaCl2, 0.081 g MgSO4 and 0.0126 g KCl with 0.2 mM 1-Phenyl-2-Thiourea aka PTU). At 2 days post-fertilization, embryos were injected subcutaneously in the perivitelline space with transfected parental and CDDP-R cells previously labeled with FAST Dil™ oil (Thermo Fisher Scientific) and treated with ± Cisplatin 5 mg/L for 3 days. Images of tumors were taken by using a fluorescent stereoscope with a K5 camera (Leica) and LAS X software v3.7.1.21655 at 100×  magnification with no binning. Images of tumors were taken right after injection (day 1) and after drug treatment (day 4). Images were automatically analyzed by using the HuginMunin software v2.7.0.0 (BioReperia AB). Tumor growth regression was calculated by dividing the number of tumor pixels on day 4 by the number of tumor pixels on day 1 in the same embryo and multiplied by 100.

To measure anatomic nerve regeneration, the following two methods were used: (1) images of regenerated spinal nerves after 21 d of SNL surgery were obtained after the dissection of perfused rats, using a ZOOM-90 6.3 megapixel camera inserted into the eyepiece of a dissecting microscope; and (2) FAST Dil oil (5 mg/ml in DMSO; catalog #D3899, Thermo Fisher Scientific) was injected into the paw subcutaneously using 31 gauge insulin syringes after the surgery of SNL. At the end of behavior testing, sections (13 μm) of L5 DRGs and longitudinal sections (40 μm) of proximal L5 spinal nerves were obtained to determine whether Dil had been transported back to the ligated L5 DRG. Costaining of Dil and a subtype of neuronal markers was then conducted (see immunohistochemistry details below) to show a clear Dil-positive neurons.