Figure 3.

Technique of immunodetection and cell picking. A–F: Laser-assisted cell picking of AMs after immunofluorescence labeling in formalin-fixed lungs. Indirect immunofluorescence was performed within 14 minutes, using anti-ED-1 and fluorescein isothiocyanate-coupled rabbit anti-mouse immunoglobulin. Original magnifications, ×40. A: An ED-1-immunolabeled AM is selected. B: Switching to bright-field microscopy, AM is identifiable in the given position next to the septum. C: Adjacent septum is removed by UV-laser. D: A sterile syringe needle is approximated. E: The AM profile adheres to the needle and is lifted. F: To prove the isolation process, switch-back to fluorescence microscopy is performed, demonstrating the fluorescent AM profile adherent to the needle. G and H: APAAP immunohistochemical staining of NOSII in formalin-fixed control and stimulated lung tissue. AMs are marked by arrows. Original magnifications, ×20. G: Control lung. H: LPS/IFN-γ stimulated lung.