This work was done In collaboration with Dalit Talmi-Frank Lab of Prof Irit Sagi. Department of Structural Biology
airSEM imaging satiation is combining technologies such as electron microscopy, optical imaging, fluorescence, spectroscopy and more, provides correlation and guidance towards specific ROI
Fibrillar collagen type 1 is a major component of the extracellular matrix in the lungs. It is presents in all anatomical lung structures, including airways, blood vessels, the interstitium of the lung parenchyma, and the basement membranes of epithelial and endothelial cells.
To differentiate the distribution and organization of fibrillar collagen type 1 within ECM in different anatomical structures of the lungs
Fig1: (a,b,c) Visualization of lung blood vessels (arrow head) and bronchus (asteriks) of mouse lung using AirSEM imaging. (d) Collagenous organization in large blood vessel (arrow head) and a bronchus (asteriks) in the mouse lung by collagen immunostaining. (e) DAPI staining of lung tissue.
Collectively, these images allow the comparison of different collagen type 1 organization and distribution within ECM of blood vessels, and bronchus. In addition it demonstrates the non-collagenous connective tissue in the interface of these two structures.
Importantly, the specially assigned metal stains that were applied to the tissue on top of the pre-labeled tissues did not cause reduction in fluorescent signal and allowed further observation by SEM.
Fig2: Comparative imaging of normal ECM lung scaffold using both airSEM and conventional SEM at the same magnifications show analogous images of alveolar lumina, walls and different levels of compartmentalization.
The significant advantages of airSEM over vacuum SEM is in terms of: Short, simple and fast sample preparation. The ability to merge information from different visualizing modalities on the same platform makes airSEM imaging station a highly available and user-friendly tool.