Troubleshooting is a pain, but a worthwhile one…

While running my 3rd and 4th trials for my experiment on TFEB localization in the presence of the pseudophosphatase MK-STYX, I have noticed that all my cells have died and that I will need to re-run my 3rd and 4th experiments in order to get data. The fact that my experiments have failed once again was disappointing and coming to terms with the fact that I will have to spend at least a couple more weeks before I can get my data been tough, but I have come to realize that the only way to get over this hill is to sit down and troubleshoot. I went back into my notebook and looked over every step that I performed and hypothesized what might have gone wrong during my execution of these steps. It turns out that I have forgotten a small but very important step: trypsinizing my cells. Without proper trypsinization, HeLa cells continue to adhere to the flask and are unable to be transported into a six-well plates during seeding. The cells that I have obtained from the flask without trypsinization were not live cells to begin with. After troubleshooting, I also wrote down the details into my notebook on what the cells should look like after each step of my experiment. This greatly reduced any doubts I had while re-running the 3rd trial today. The big lesson that I learned through all this is that trial-and-error is what research is all about and that disappointment and troubleshooting are also integral parts of the research progress. We should not only seek to get the data we want, but to also appreciate the lessons that we are able to learn when experiments fail.


  1. Wow Y-Nhi, I am so amazed at your open minded way of thinking. I too had a lot of problems with cells dying more multiple trials. Given that, I totally understand your frustration behind troubleshooting and optimizing procedure techniques. I am very excited to see how your last week of trials turns out. Good luck!

  2. kmreed01 says:

    Hi Y-Nhi! I’m so sorry to hear that your most recent trials didn’t work out. I’ve been having some trouble with cells dying, too, so I totally understand your frustration. I’m glad to hear that you’re keeping a positive attitude and that you identified the issue– research is definitely all about learning from your mistakes. Hope your last few trials of the summer go well!

  3. Wow! It’s very interesting to see how people’s research actually does fail sometimes, but it is a very worthwhile task to stop and backtrack and see what you have may done wrong and then re-work the experiment. It also goes to show that sometimes even the smallest details can cause results to be spewed in a completely different direction than what you are hypothesizing or wanting to happen. I am happy that you found that small detail and that you can now go back and do better experimentation. I am also very impressed by the different methods that you used in order to make the experiment work in a different way and that you could possibly get the results that you have been looking for in the long run.