Cell Labeling on Si
Below is an image of HUVEC grown on RTP treated W303 for 72 hours. The cells were seeded at high density and therefore reached confluence prior to day 3. The first row shows the membrane area of the sample; the membrane is not intact. The second row shows the edge of the sample. Live and dead staining was conducted using the live/dead assay (calcein AM and ethidium homodimer). The images were taken with the well side of the sample facing upward.
It is clear that the live/dead assay can be used to visualize the cells on the membrane. This is the way we will be imaging the cells for the growth experiments. The “dead” assay will also be useful in comparing the health of the cells as they proliferate on different surfaces.
Going forward in planning the growth experiment I have a few questions:
1. Some of the bright spots in the “dead cell” image seem to be small, are these all cellular fragments? If so, can we correlate these images to a number of dead cells in the future? I would not expect one cell to produce one fragment.
2. Also, do we think we could distinguish between necrotic and apoptotic death using this assay?
