High resolution co-culture images

One of the points I’d like to discuss in my co-culture paper is the advantageous imaging qualities of pnc-Si compared to ‘translucent’ commercial transwell membranes.  This has been evident in many of my posts of Live/Dead stained cells, even at 20X.  I thought a higher resolution (100X) image would emphasize this point even more since the 100X objective has a much shorter working distance than the 20X.  Here, I show images of bEnd3 endothelial and NG101815 glial cells on opposite sides of transwell membranes (with bEnd3 facing the objective) observed at 100X with DIC.

In the top row, I’ve focused at the endothelial focal plane in the left image and the glial focal plane in the right image.  Endothelial cells are barely discernible because they are so flat.  In general, resolution through this polymer membrane isn’t that great and you certainly can’t see both cell types in the same image.  In the bottom row, I’ve included 2 different ROI for pnc-Si.  Here you can clearly see both cell types and they are both in focus (even endothelial vacuoles are visible).  I think this side-by side comparison make the point that pnc-Si allows imaging of co-cultures in transwell devices with a resolution not attainable with commercial devices.