Development of Real-time Subcellular Dynamic Multicolor Imaging of Cancer-Cell Trafficking in Live Mice with a Variable-Magnification Whole-Mouse Imaging System

Abstract

With the use of dual-color fluorescent cells and a highly sensitive whole-mouse imaging system with both macro-optics and micro-optics, we report here the development of subcellular real-time imaging of cancer cell trafficking in live mice. To observe cytoplasmic and nuclear dynamics in the living mouse, tumor cells were labeled in the nucleus with green fluorescent protein and with red fluorescent protein in the cytoplasm. Dual-color cancer cells were injected by a vascular route in an abdominal skin flap in nude mice. The mice were imaged with an Olympus OV100 whole-mouse imaging system with a sensitive CCD camera and five objective lenses, parcentered and parfocal, enabling imaging from macrocellular to subcellular. We observed the nuclear and cytoplasmic behavior of cancer cells in real time in blood vessels as they moved by various means or adhered to the vessel surface in the abdominal skin flap. During extravasation, real-time dualcolor imaging showed that cytoplasmic processes of the cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Both cytoplasm and nuclei underwent deformation during extravasation. Different cancer cell lines seemed to strongly vary in their ability to extravasate. With the dual-color cancer cells and the highly sensitive whole-mouse imaging system described here, the subcellular dynamics of cancer metastasis can now be observed in live mice in real time. This imaging technology will enable further understanding of the critical steps of metastasis and provide visible targets for antimetastasis drug development. (Cancer Res 2006; 66(8): 4208-14)

Fluorescent LYVE-1 Antibody to Image Dynamically Lymphatic Trafficking of Cancer Cells In Vivo

“…Tumor cell trafficking through mouse lymphatics imaged with AlexaFluor-conjugated LYVE-1 was imaged in vivo in real time using live-mouse fluorescent imaging technology previously developed in our laboratory [13][14][15]. After in vivo staining of mouse lymphatic endothelium with AlexaFluor-conjugated LYVE-1, human pancreatic tumor cells expressing red-fluorescent protein (RFP) were injected into the inguinal lymph node and surrounding tissues and lymphatic trafficking was imaged.…”

Spatial–temporal FUCCI imaging of each cell in a tumor demonstrates locational dependence of cell cycle dynamics and chemoresponsiveness

“…A skin-flap abdominal imaging window [11][12][13] was used to longitudinally visualize the cell cycle phase of cancer cells within tumors using FUCCI and confocal laser scanning microscopy (CLSM) (Fig. 1A).…”

Time-Course Imaging of Therapeutic Functional Tumor Vascular Normalization by Antiangiogenic Agents

“…The Olympus OV100 (Olympus Corporation) is equipped with GFP, RFP, and near-infrared (NIR) fluorescence filter sets enabling multichannel coimaging (16,17). For conducting dual-channel imaging with paclitaxel-BODIPY 564/570 or with the NIR blood pool agent AngioSense 750 , the probes were injected via the tail vein 4 hours before imaging (20 and 2 nmol/mouse, respectively).…”

“…For conducting dual-channel imaging with paclitaxel-BODIPY 564/570 or with the NIR blood pool agent AngioSense 750 , the probes were injected via the tail vein 4 hours before imaging (20 and 2 nmol/mouse, respectively). Eight-bit planer images were acquired through the RFP and 750-nm channels of the OV100 similar to what we previously described (16). GFP Z-stacks were obtained between 2 and 4 mm at 10 mm steps at the Â1.6 zoom level.…”