Lungs were fixed by inflation with 10% buffered formalin under 20 cmH2O of pressure, paraffin imbedded, and stained with PAS, Mason-trichrome stain, or subjected to immunohistochemistry (IHC) or immune-fluorescence (IF) staining using standard techniques28 . PAS, trichrome or IHC-stained slides were scanned at 20x using a Nanozoomer HT imaging system (Hamamatsu) as above. Positively stained pixels were counted across the entire section using ImageScope Software and the “Positive Pixel v9” algorithm as previously described9 . For immunofluorescence experiments we utilized antibodies directed against Mac3 (Ebioscience) and cleaved caspase 3 (Cell Signaling) and Alexa Fluor-conjugated secondary antibodies (Invitrogen). Stained slides were visualized using an Olympus BX51 microscope and a Retiga-2000R camera system (QImaging).
Nanozoomer ht imaging system
Manufactured by Hamamatsu Photonics
The Nanozoomer HT is a high-throughput digital slide scanning system designed for whole slide imaging. It captures high-resolution images of microscope slides and converts them into digital files for analysis and storage.
Automatically generated - may contain errors
2 protocols using nanozoomer ht imaging system
1
Lung Fixation, Staining, and Imaging
2
Lung Fixation, Staining, and Imaging
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