Recovering From Mistakes in the Lab

Today I spent nearly three hours on the LSM980 confocal microscope collecting images. After finishing the imaging session, I felt relieved and happy because the experiment itself went smoothly. The final step seemed simple: replace the HEPES buffer with 2.5% glutaraldehyde (GA), then seal the dishes with parafilm for fixation.

But then I made a mistake.

I removed the lids from all the dishes at once while changing the buffer. Afterward, I suddenly realized that I no longer knew which lid belonged to which dish. Since the dishes contained different experimental conditions, mixing them up would have made the entire experiment unusable.

Fortunately, before using the confocal microscope, I had collected overview images (4k and 10x) of each dish and specific square on the Keyence microscope. I went back to the Keyence images and carefully compared the cell distributions and patterns dish by dish. It took another hour, but eventually I successfully mapped every dish back to its correct condition.

In the future, I should label the bottom of each dish instead of relying on the lids.