Bad imaging day

This week I finally went live, i.e. started live imaging.

The first session, on Tuesday, was ok.

Then today…

I came early to start up the systems. The most important thing is the heating chamber. Heating is important since the temperature can affect the quality of the imaging. How? The metal tray (where you place your sample) expands when heated. The lenses in the objective also. The immersion oil changes viscosity. So does the cell media. So when imaging mammalian cells at 37C, it takes time for all components to equalize their temp on 37C.

3 hours later, I was ready to image. I started the camera, the computer, the lasers. The only thing left to do before starting is align the laser so it will go directly up through the objective (at 90 degree to the sample). The laser sometimes drifts, or users change it and so one has to check before starting.

The laser was waaayyy off the target. I tried my best. Then, over the next  4-5 hours(!), three other people tried until our physicists succeeded, in a way (it’s centered but the laser looked dispersed, not sharp).

Problem is, the temp in the chamber isn’t equalized now (since we kept opening the chamber, switching objectives, removed the metal plate holder etc…). I tried imaging without waiting, but the z-axis kept drifting. So now I have to wait another half-hour or hour to let everything heat up properly and it’s already 6pm.

Guess I won’t be doing lots of imaging today…

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