Home › Cell Biology › Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma
Cell Biology JoVE (Open Access) Citable · DOI

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

DOI: 10.3791/2941-v
What you'll learn
  • ✓ Construct LifeAct-mCherry labeled head and neck cells
  • ✓ Perform orthotopic xenograft injection into mouse tongue
  • ✓ Prepare tongue tissue for ex-vivo imaging
  • ✓ Analyze tumor invasion using two-photon microscopy
Protocol

Biopharma Insights A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Difficulty
advanced
Total time
~2 weeks for cell line generation, ~2 hrs for imaging analysis
Model organism
Mouse (orthotopic xenograft)
Biosafety
BSL-2

Steps

1
Generate lentiviral vectors encoding LifeAct-mCherry

Prepare lentiviral constructs for stable expression of LifeAct-mCherry in head and neck cancer cell lines.

▶ 01:46
2
Produce stable LifeAct-mCherry expressing cells

Transduce head and neck cell lines with lentivirus and select for stable LifeAct-mCherry expression.

▶ 03:19
3
Inject cells into mouse tongue orthotopically

Perform orthotopic xenograft formation by injecting labeled cancer cells into the mouse tongue.

▶ 04:30
4
Prepare mouse tongues for ex-vivo imaging

Harvest and mount tongue tissue from tumor-bearing mice for ex-vivo multi-photon imaging.

▶ 05:56
5
Image tongue tumors and analyze invasion

Use two-photon microscopy to capture images of labeled tumor cells, then analyze invasion patterns.

▶ 06:27
6
Review representative tumor images

Examine example images from the imaging and analysis steps to illustrate tumor invasion into tongue tissue.

▶ 09:55
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