India Inktinflatie: een kleuringsmethode om tumorknobbeltjes te visualiseren

Begin by placing a euthanized mouse ventral side up on a surgical bed and securing the limbs of the animal. Using scissors, cut along the midline through the rib cage up to the salivary glands. Place a pipette tip underneath the trachea to elevate and isolate it. Rotate the surgical bed 180 degrees. Insert a syringe needle containing an India ink stain into the trachea and inject the ink into the lungs until you feel resistance.

Cancer cells secrete a large amount of lactic acid that generates a strong negative charge on the cell surface. When these cells are stained with India ink, an acidic stain, the cell surface repel the stain and remain unstained.

Use scissors to cut the trachea and slide a pair of tweezers under the lungs. Gently pull the lungs out of the animal's body. Rinse the lungs in water and transfer them into a vial containing Fekete's solution to remove excess stain and fix the tissue in a fume hood. The tumor nodules appear white against the dark background.

In the following protocol, we will perform the India ink inflation technique to validate lung metastasis.

To validate the metastasis of the primary tumor to the lung, first place the mouse in the supine positio