Tumour burden of Molm-13 and MDS-L in zebrafish larvae with or without anti-cancer treatment. Molm-13 and MDS-L cells were stained with CellTracker™ Deep-Red and 4 nL of a 10 106 cells·ml−1 suspension engrafted into zebrafish larvae 2 dpf by injection into the posterior cardinal vein. Following engraftment, the larvae were imaged daily using spinning disk confocal microscopy. Treatment of Molm-13 cell-xenografted larvae consisted of a single 4 nL 1 mM daunorubicin injection at the day of the transplant, whereas MDS-L engrafted larvae were given daily injections of 4 nL 1 mM azacitidine. Control samples consisted of transplanted larvae without injection as well as larvae injected with 4 nL Milli-Q® water at the day of transplantation for Molm-13 and daily for MDS-L. Imaging and treatment were continued until the larvae reached 5 dpf. Using our software tool, fluorescent cells were counted and segmented based on the confocal images. The total volume of all segmented objects in larvae engrafted with Molm-13 and MDS-L are shown in A and C, respectively. Using the volume threshold of 1000 µm3 determined from the images in Fig. 2C and D, the filtered total cell volume was determined (B and D). n=15 except for MQ-injected larvae, where n=6. Significance between injected larvae and non-injected controls were found using two-tailed Welch's t-test. Not annotated: P>0.05, *P≤0.05, **P≤0.01.
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