Results

Fluorescence complementation

Results

We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!

Figure 1. Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.

Figure 2. Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.

Figure 3. Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.

Figure 4. Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.

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