Home › Cell Biology › Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models
Cell Biology JoVE (Open Access) Citable · DOI

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

DOI: 10.3791/57088-v
What you'll learn
  • ✓ Engineer cancer cells expressing NIS-FP dual reporter for PET/fluorescence imaging
  • ✓ Perform [18F]BF4-PET/CT imaging to track tumor progression in rodent models
  • ✓ Validate reporter-specific uptake via ex vivo analyses and autoradiography
Protocol

We describe a protocol for preclinical in vivo tracking of cancer metastasis. It is based on a radionuclide-fluorescence reporter combining the sodium iodide symporter, detected by non-invasive [18F]tetrafluoroborate-PET, and a fluorescent protein for streamlined ex vivo confirmation. The method is applicable for preclinical in vivo cell tracking beyond tumor biology.

Difficulty
advanced
Total time
~7–10 days (cell engineering 3–4 days; tracer synthesis 4–6 hrs; imaging 2–4 hrs per animal; ex vivo analysis 1–2 days)
Model organism
Rodent tumor xenograft models; cancer cell lines (HEK293-derived or similar)
Biosafety
BSL-2

Steps

1
Engineer cancer cells expressing NIS-FP reporter

Transduce or transfect cancer cells with the sodium iodide symporter-fluorescent protein fusion construct. Characterize expression via flow cytometry and fluorescence microscopy to confirm stable dual-modality reporter expression.

▶ 01:40
2
Synthesize [18F]tetrafluoroborate radiotracer

Produce [18F]BF4 via cyclotron or generator-based synthesis and purify using standard radiochemical methods. Verify radiochemical purity and specific activity before imaging experiments.

▶ 01:40
3
Perform nanoPET/CT imaging of tumor-bearing rodents

Inject [18F]BF4 tracer into tumor-bearing mice or rats, allow uptake kinetics to occur, then acquire PET/CT images on a preclinical scanner. Reconstruct and analyze images for biodistribution and tumor uptake.

▶ 04:04
4
Perform ex vivo tissue analysis and autoradiography

Harvest tumors and organs post-imaging, measure radioactivity via gamma counter, and prepare tissue sections for autoradiography. Correlate PET signal with fluorescence microscopy to confirm reporter specificity.

▶ 05:40
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