In the normal situation, analyzing 9 mice is not too big a deal. However, this is my first time screening, so to be more careful, we collected 5 different tissues including bone marrow, spleen, payer's patch, peritoneal cells and thymus. Therefore, yes, I have 45 cells to count, which took me almost 3 hours!
I know I am not phisically strong enough to work so long right now, but this is a very important experiment and there's no way I can stop and come back to work the next day. When I finished counting cells and started to stain with antibodies, it was almost 6 pm. I was exhausted. And here comes the tragedy. After I washed the cells and centrifuge them, cells were frozen!!!!!
I checked the centrifuge, the temperature setting was at -2 C. It is very wired because I was the only one using the machine yesterday, and I didn't adjust the setting at all. It is possible that somebody accidently touched it and wasn't aware of, and of course, I might be the "somebody". Anyways I was shocked for couple seconds and then immediately put the cells into 37C waterbath trying to rescue them. But I know once cells are frozen, they might all be lyzed. I sadly kept staining and fixed the cells. When I left the lab, it was almost 9pm.
It is a very bad idea to do a very long experiment alone. Although my boss helped me to collect the tissues in the morning, it is still very tiring. I was sad and frustrating this morning when I told my boss what happened this morning in the lab meeting. Other people in the lab told me the centrifuge is old and the temperature setting sometimes just goes crazy. Oh well, I guess I am just unlucky, the machine knows I am new to the lab.
Since cells are still there, I analyzed them by flow cytometry this afternoon. Surprisingly, they looked OK!
Maybe I am not that unlucky afterall. Haha.Conclusion: Never ever do any experiment alone during after hour!!!!!!
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