Figure 5

Confocal and multiphoton imaging of Carmine Alum fluorescence in fixed mammary gland whole mounts. Confocal and bright field imaging of TEBs close to the surface of a mammary gland reveals the morphology and arrangement of TEBs relative to the plane of the coverslip. A. Imaging of a 10 week mammary gland from an EGFP mouse was performed using a 543 nm HeNe laser and conventional confocal imaging of single XY planes since the TEB was close to the surface of the gland. A typical filter set for imaging of red dyes was used (emission bandpass of 565–615 nm). B. Bright field imaging of the same TEB was not very informative. C. Multiphoton imaging of an EGFP-mouse mammary gland was performed using a Chameleon XR Ti-Sapphire laser tuned to 750 nm (2.1%), and PMT Ch3 was used with bandpass filter 565–615 and pinhole set open to 1000 μm. Scale bars = 50 μm except for the inset indicated by a white box, 20 μm.