Journal: Communications Biology
Article Title: Tumour-on-chip microfluidic platform for assessment of drug pharmacokinetics and treatment response
doi: 10.1038/s42003-021-02526-y
Figure Lengend Snippet: Spheroids were treated in the chip with SN38 and AZD0156 at different schedules using the microfluidic setup. Spheroids were recovered from the chip at day 7 and DNA double strand break damage was assessed via the presence of γH2AX. Scalebar = 100 µm. a Representative images of nuclei (blue/Hoechst 33342) and γH2AX (green), b quantification of γH2AX positive cells. (A-SN38, B-SN38 + AZD0156 3/7 with 24 h gap, C-SN38 + AZD0156 3/7 with 72 h gap, D-SN38 + AZD0156 1/7, E-SN38 + AZD0156 7/7, F-control). Cleaved caspase 3 (CC-3) was used to quantified apoptotic cell death ( c ), and Ki67 to measure the effect on proliferation ( d ). N ≥ 5 per condition from two independent experiments. Box-plots show median (centre line); box limits are 25th to 75th percentile; whiskers represent min and max values. Statistical analysis was carried out using 1-way ANOVA, Tukey’s multiple comparisons, CI = 95%, **** p < 0.0001; *** p < 0.001; ** p < 0.01; * p < 0.1.
Article Snippet: Then, triple labelling was performed overnight, in static conditions, using primary conjugated antibodies for γH2Ax (gamma H2AX [p Ser139] Antibody [Alexa Fluor 488], NB100-384AF488, Novus Bio/BioTechne), caspase-3 (CC3, Caspase-3 Antibody (31A1067) [Alexa Fluor 594], NB100-56708AF594, and KI67 (Anti-Ki67 antibody [EPR3610] (Alexa Fluor 647), ab196907, Abcam) at 1:500 dilution each.
Techniques: