Fig. 3: Barcoded population frequencies and metastatic burden of MDA-MB-231 cells in primary tumours and lung tissue.
From: LeGO-3D: 3D imaging of lung metastases and vascularisation using light sheet fluorescence microscopy
a Matched primary tumour, b confocal lung lobe section and c light sheet lung lobe. Scale bar 1000 μm. a–c Magnified regions from white boxes. Scale bar a 90 μm, b, c 180 μm. Arrows indicate different BPs. d Stacked bar chart shows biomass of seven BPs in injected cells, primary tumour and lung from one animal. BP biomasses were determined by flow cytometry (injected cells) or from imaging data (tumour, lung). Lungs from (e–i) intravenously (IV) injected and (j–m) mammary fat pad (MFP) injected mice. Scale bar 1000 μm. n Stacked bar chart shows contribution of seven BPs to metastasis biomass in IV and MFP lungs. BP biomasses were determined from imaging data. d, n Number of BPs is shown above each bar. Key is shown below stacked bar chart, where each colour represents a unique BP. Number of metastases with volume o 100,000 μm3 to 1,000,000 μm3 (100–1000 cells, small), p 1,000,000 μm3 to 10,000,000 μm3 (1000–10,000 cells, medium), q 10,000,000 μm3 to 50,000,000 μm3 (10,000–50,000 cells, large), r above 50,000,000 μm3 (>50,000 cells, very large) in IV and MFP mice. Independent t-test, P* = 0.030645. s Number of metastases normalised to lung volume in IV and MFP mice. t Percentage metastasis volumes to lung volumes in IV and MFP mice. u Proportion of monochromatic and polychromatic metastases in IV and MFP mice. Paired t-test, P**** = 0.000043, P*** = 0.000450. v Percentage metastasis volumes to lung volumes of monochromatic and polychromatic metastases in IV and MFP mice. All box plots show the median and interquartile range (IQR), with lines representing 1.5*IQR. n = 5 mice for IV and n = 4 mice for MFP from 4 independent experiments. The number of monochromatic metastases analysed for IV-injected mice ranged from 384 to 5300, and for MFP-injected mice ranged from 1057 to 3196. The number of polychromatic metastases analysed for IV-injected mice ranged from 21 to 419, and for MFP-injected mice ranged from 58 to 219. In o-r, subsets of metastases were analysed based on metastasis volume. For one experiment, mice from the IV and MFP groups were injected in parallel (highlighted by the burgundy-coloured dots).
