Interest in ultrasound perfusion imaging has grown with the development of more sensitive algorithms to detect slow blood flow. Unfortunately, there are not many phantoms that can be used to evaluate these techniques. Some have used small linear tubes, while others have adapted dialysis cartridges. Here we propose a technique using conventional gelatin cast around a sacrificial polymer network. Specifically, we form a gelatin phantom, doped with graphite scatterers to mimic the diffuse scattering in soft tissue, around a polymer resin. The resin structure can be dissolved leaving behind a network of small randomly oriented channels that are connected to a large channel which is connected to a pump to perfuse blood mimicking fluid through the phantom. The phantoms were qualitatively demonstrated to show perfusion through visual confirmation and the speckle SNR, and speed of sound were calculated.
We report the use of antibody-conjugated quantum dots (QDs) to monitor the expression dynamics of the membrane bound cytokine receptor interleukin-2 receptor-α (IL-2Rα) throughout the course of Jurkat T cell activation. Maximal receptor expression is observed 32-48 hours after activation, followed by a sharp decrease subsequent to 48 hours consistent with IL-2R internalization. Fluorescence microscopy, ELISA, and FACS analyses were used to verify controlled activation and specificity of QD labeling. Additionally, confocal microscopy demonstrated receptor internalization subsequent to expression and QD labeling. Antibody-conjugated QDs provide a convenient means to rapidly determine cell state and interrogate end products of cell signaling pathways. Interrogation of other signaling pathways can eventually be carried out in a similar manner upon identification of relevant membrane associated receptors. Ultimately, the multiplexing capabilities of QDs will allow the examination of several signaling pathways simultaneously and aid in toxin detection and discrimination.
Vascular endothelial growth factor (VEGF) is one of the most potent mediators of both physiologic and pathologic angiogenesis. Normal physiologic induction of VEGF occurs during periods of extreme growth, wound healing, as well as immune inflammatory response. Pathologically, however, VEGF is largely responsible for tumor induced angiogenesis and cell survival. Traditional methods of VEGF expression analysis involve either in vitro studies, or highly invasive in vivo methods. We have developed a unique transgenic mouse model (VGL) that possesses a truncated human VEGF promoter attached to a GFP-Luciferase fusion protein. Incorporating this model with both spontaneous and orthotopically injected tumors allow VEGF promoter activity to be visualized in vivo by luciferase luminescence in response to tumor growth non-invasively and over time. By also utilizing bioluminescent tumor cells, we were able to generate models that identify host, tumor, or combined VEGF promoter activity. Results indicate that tumor tissue is responsible for the majority of VEGF promoter activity during tumor growth. Additional studies into the mechanism by which tumor cells initiate VEGF production will yield much needed insight into tumor survival. In conclusion, we have shown that the VGL bioluminescent mouse model is indeed capable of yielding compelling information on host-tumor interactions.