HA-1077 ROCK inhibitor implanted sc in the shoulder of a female nude athymic

Local institutional rules for animal HA-1077 ROCK inhibitor experimentation. About 8106 B-clone and MCF7 cells MCF7 parental 5106 in 40 ml PBS mice were separately suspended in 50 ml of Matrigel and implanted sc in the shoulder of a female nude athymic M. For each mouse, clone B cells were on the right or C T inoculated left, and MCF7 parental al Ty, from estradiol valerate 17 pellet was sc into the neck region of implanted support tumor growth. Tumors lie to grow for 5 to 10 mm in diameter before the Mice were subjected to imaging. The tumor size was en monitored on days imaging caliper measures the diameter L gr eren longitudinal and transverse directions, the tumor volume was then calculated using the formula: Tumor volume 1/2. Mice in eight M Microwave ultrasound was performed to obtain accurate measurements of tumor volume. The tumors were measured in orthogonal directions 3-4 times until 3 days imaging and tumor volume is then calculated using the formula: tumor volume pxyz / 6 Fluorescence imaging optical mouse were incubated with 2% isoflurane in oxygen 2 L / and on a heating pad. The Mice were anesthetized w Held during the fluorescence imaging with a time domain in vivo fluorescence imaging of small animals. Diode laser at 670 nm was used for pulsed excitation. The average power of the laser is maintained at about 1 mW at a repetition frequency of 80 MHz. The width at half maximum of the laser pulse was about 100 ps. A 693 nm long pass filter was used to is the signal emitted by fluorescence. The eXplore Optix detector is a photomultiplier with a fast time correlated Einzelphotonenz Hlung system for time resolved Coupled ste measurements. The general Zeitaufl Solution of the detection module was about 250 ps. Before the injection of fluorescently labeled Affibody, images were acquired prior to injection, the autofluorescence background signal or sp To remove ter.
The Mice Were injected iv with 500 pmol Caspase 9 Affibody AlexaFluor680 in PBS in a total volume of 150 ml of gel St. The images were recorded after injection after 5 hours on the basis of pilot data. The Mice were positioned on the c Tees and to the right one after each time adjusting the H theirleft Height of the table to ensure optimal fluorescence imaging of each tumor adjusted to erm. The field of view for each sample was adjusted so that the entire tumor was imaged with a surrounding tissue. The scan resolution and high 1.0 to 1.0 mm. The acquisition time varies between 3 and 9 minutes per scan. The images were analyzed by the OptiView. First, the pre-injection image for background autofluorescence signal was removed. Then, a region of interest around the tumor was drawn. Average standardized figures were calculated for each ROI. In pilot studies, the coefficient of variation for the average normalized Z Hlungen determined. In six imaging sessions were clone B xenografts imaged 3 times for about 5 hours after a single injection of Affibody. For each analysis, the Mice positioned, the imaging system is recalibrated, and the means of the Z Hlungen were normalized F Is independent Dependent. Imaging procedures were initiated when tumors were about 5 to 10 mm. Rst Were the pre-recorded images. Then Mice were divided into 2 groups.