Digital microphotography system dfx1200

Lungs were fixed by inflation with 1 mL of 10% neutral buffered formalin then excised and immersed in neutral buffered formalin prior to routine paraffin embedding. 5-μm sections were cut and stained with H&E and mucicarmine. Sections were analyzed by light microscopy and microphotographs were taken using Digital Microphotography system DFX1200 with ACT-1 software (Nikon, Tokyo, Japan).

Mice were sacrificed according to approved protocols, and the lungs were immediately perfused with sterile PBS by transcardial perfusion through the right ventricle. The pericardium and trachea were exposed by dissection, and an incision was made in the trachea for the insertion of a sterile flexible cannula attached to a 3-ml syringe. The lungs were slowly inflated with 0.5-0.7 ml 10% ultrapure formaldehyde (Polysciences, Inc., Warrington, PA), excised and immediately fixed in 10% ultrapure formaldehyde for 24-48 hours. The lungs were then transferred to 70% ethanol and subsequently mounted into cassettes and paraffin embedded by personnel at The University of Texas Health Science Center at San Antonio Histology and Immunohistochemistry Laboratory. After paraffin embedding, 5-mm sections were cut and stained using hematoxylin-eosin (H&E) with mucicarmine at McClinchey Histology Labs, Stockbridge, MI. Sections were analyzed with light microscopy using a Nikon microscope and microphotographs taken using Digital Microphotography system DFX1200 with ACT-1 software (Nikon Co, Tokyo, Japan) at the Ann Arbor VA Health System.