Concurrent TEER and Phase Microscopy with Yuchen Wang

For the first time, we did time-lapse phase microscopy (1 frame / 5 seconds) looking at endothelial cells through the ITO in the static BBB device (device A bottom left well) and measuring the TEER concurrently. Here’s the video, just shy of 15 minutes of recording.  (reporting a low resistance: 5-10 ohms, shorted).

TEER Test 15min

We started the TEER measurement at frame 30, left it running continuously. The device was bumped around frame 100. We then did another video (1 frame/ 10 seconds) on another well (top right well, device A). Here’s the video, 30 minutes of recording. Again, the EVOM2 TEER  tool was left continuously running  after frame 15. I apologize for the quality; something happened in the encoding process (the raw avi doesn’t have the blue/green artifact). If brighter images are desired, we can adjust the exposure time to compensate. We seem to be limited to 177 frames we can capture on phase microscope PC.

TEER Test 30min

If you stare at the videos long enough, you can see some endothelial cells moving around and some possible bacteria floating around (contamination = not good). It’s been 4 days since the cells were seeded on the device. Looking at our previous images, we were concerned that the TEER measurement was harmful to the cells in some way, causing them to ball up and die. It’s not clear from these images that all the cells are experiencing something harmful. The control to this experiment would be to leave the cells alone for the same amount of time and observe. We also hypothesize that leaving the cells out of the incubator for extended periods of time (1hr here) may not be the best because of non-optimal temperatures and gas concentrations.

It’s very nice to have transparent electrodes to see all the little details unfold 🙂