A 3D reconstruction of the cardiac outflow tract (OFT) region (horizontal and vertical rotation) from stage-14 wild-type embryo stained for Tinman (Tin; red) and Msp300 (green). Tin staining reveals the cardiac cells and, located ventrally to cardiac outflow muscles (COM), cvr cells (red cells without Msp300 staining). The two anterior cardioblasts coexpressing Tin and Msp300 (ACB cells) are seen associated with COM (refer to Fig. 1). Notice the position of ACBs linking the two COMs (frontal view) and located dorsally with respect to COM fibers (lateral view). Movie is SI Movie 2 in Proc Natl Acad Sci USA (2008) 105: 2475-2480. Movies from this publication include CIL# 25536, 25537, 25538, 25539, 25540, 25541.
Stage 14 embryos were stained with primary antibodies: rabbit anti-Tim, guinea pig anti-Msp300. Species-specific secondary antibodies were conjugated to Alexa Fluor 488, Cy3 or Cy5. The double stained embryos were visualized using a Zeiss LSM 510 Meta confocal microscope. Images representing 0.5 mm optical sections were compiled and processed by Volocity software (Improvision).
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