Ultrasound imaging.

<p>A. The mice were anesthetized with isoflurane and mounted on the heated imaging table with continuous monitoring of vital signs. After visualization of the bladder with the Vevo 700® small animal imaging platform the skin was perforated with a 30G needle. B. Ultrasound visualisation of normal mouse bladder in sagittal section with typical dimensions indicated (lumen dimensions 4.4×6.5 mm; wall thickness 0.25 mm).</p

Inoculation of tumor cells.

<p>A. Detection of the needle on the ultrasound screen. B. Perforation of the skin and abdominal wall muscles. C. Needle insertion into the bladder wall without penetration of the mucosa. D. Injection of PBS (50 µl) between the muscular layer and the mucosa. E. Guidance of second needle into the artificially created space. F. Injection of tumor cells suspended in Matrigel®.</p

Longitudinal imaging of xenograft growth.

<p>Tumor growth was measured at regular time intervals by: A. bioluminescence imaging, and B. ultrasound. C. Correlation of bioluminescence and xenograft volume for all three cell lines. D. H&E section of a representative UM-UC1 luc xenograft demonstrating invasive growth into the muscle (*) without invasion into adjacent organs. All tumors originated from the anterior bladder wall and often occupied most of the bladder lumen without infiltrating the posterior wall (**).</p

Ultrasound-guided intratumoral injection of treatment agents.

<p>A. The xenografts were visualized by ultrasound and either VSV (1.05×10⁷ pfu) dissolved in 25 µl PBS or PBS alone was injected through a 30G needle into the center of the tumor. B. 48 h after injection of VSV, all xenograft tumors showed positive staining for VSV-G around the injection site which correlated to TUNEL staining. C. VSV-G and TUNEL staining were negative after PBS injection alone.</p

Percutaneous tumor cell injection – procedure, complications and growth.

<p>Percutaneous tumor cell injection – procedure, complications and growth.</p

Treatment of xenograft tumors by chemotherapy.

<p>A. Mice bearing UM-UC13 luc tumors showed a remarkable decrease in tumor volume after systemic therapy with a combination of gemcitabine and cisplatin starting on day #28 after inoculation, compared to PBS control (** = P<0.01). B. Xenograft perfusion was measured by injection and ultrasound imaging of non-targeted microbubbles in UM-UC13 luc xenografts before and 5 days after administration of control agent (PBS; left panel) or systemic chemotherapy (gemcitabine/cisplatin; right panel). Perfusion was quantified as contrast percent area. Representative single results out of 4 measured animals per group are shown.</p

Intramural injection of agarose.

<p>The sagittal H&E section of a whole murine bladder demonstrates a layer of gel strictly in the lamina propria between mucosa and the muscularis propria.</p