In order to study melanoma lung metastasis in aged hosts, we did RO injection of B16F10Luc2 melanoma cells (expressing firefly luciferase) into both young (16 weeks) and old (>2 years) mice. However, we noticed a fair amount of signals were coming from the eye area when doing IVIS on these mice meaning many of the cells injected were trapped in the eye and forming localized tumors instead of metastasizing to the lung. I wonder if this is a normal thing and if there are any ways to prevent this from happening.
Retro-orbital inoculation of tumor cells is very likely to give rise to trapping of some of the injected tumor cells in the eyebed and hence tumor formation. It is better to introduce such cells via the tail vein. If a second population of cells needs to be injected i.v. as well (for example primary lymphocytes) these could be injected retro-orbitally (provided this is still allowed under the pertinent legislation). These should be primary cells in rder to avoid the problem raised in the question. It should be mentioned that detection of tumor cells in the lung upon i.v. injection is not a very good model for metastasis, its appearance most likely representing trapping of the tumor cells in the vasculature of the lung. It is a valid model to measure, for example, the potency of anti-tumor immunotherapy approaches.
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