Sp8 405 488 561 633 laser confocal microscope

Pregnant C57BL/6 damns were injected at E14.5 with 10μg fluorescein-tagged pIC (InvivoGen) supplemented with untagged pIC to a final dose of 20mg/kg. Mice were sacrificed 24-h post injection and whole fetuses were placed in tissue molds, suspended over 2-methylbutane (Fisher Scientific), and imbedded in O.C.T. compound (Fisher Scientific). Embryos were frozen over dry ice and stored at −80°C. 20μm cryosections were collected at −20°C to −17°C and placed onto slides. Slides were allowed to dry in the dark at room temperature (RT) for 30 min prior to fixation with 10% formalin at RT for 15 min in the dark. Sections were washed 3X with 1X PBS^−/− and dried overnight protected from light. Sections were stained with DAPI and coverslip was applied and secured before imaging. Fetal liver images were captured at 63X magnification using a Leica SP8 405-488-561-633 laser confocal microscope.