Imaging Circulating Tumor Cells in Freely Moving Awake Small Animals Using a Miniaturized Intravital Microscope
Figure 1
Experimental mouse metastatic breast cancer model.
(A) Schematic of lentiviral construct comprising a fusion reporter gene (Luciferase-2 and enhanced GFP) under the control of the ubiquitin promoter, used to establish the imageable metastatic mammary carcinoma cell line 4T1-GL. (B) FACs analysis of GFP fluorescence, comparing the stable cell line 4T1-GL at passage 2 and passage 12 (resp. P2 and P12) to wild-type 4T1 cells (4T1-WT). (C) Metastatic tumor growth in the lungs as monitored non-invasively by Bioluminescence (BLI) imaging, following a systemic injection of 1×106 4T1-GL cells via the tail vein (n = 7). (D) Biodistribution of metastatic cells, 12 days after systemic injection (n = 7) in the following organs: Lungs, Liver, Heart, Kidneys Spleen, Bone marrow, and corresponding quantification of BLI signal per organ (n = 7). (E) CTCs in 100 µL blood samples of mice (n = 7) at various times from day 0 (immediately after injection) to 12 days after injection and corresponding signal quantification. Positive BLI signals correspond to <20 CTCs/100 uL of blood.