Rising above setbacks in science

Not everything works in research. And that is okay. But why we do often feel that is not?

This post will explore the 'darker side' of science...when things don't work. I hope sharing some of my experiences and thoughts can help those of you struggling with setbacks that you may be facing.

When research just doesn't work

The first thing I would like to share is something that happens to all of us...when your experiments just don't work. How frustrating is that? Let me share a story back when I was a graduate student...

My graduate work was in cell biology and for this project, I was investigating the activation of a protein called p70S6k. While there a number of ways to detect activation, one of the simplest ways to do so is by detecting an upward shift in its position when it is run through a gel (western blot analysis; I won't go into detail why as many of my readers are from different fields, and it is not relevant to the key part of the story).

My research was going great...I was routinely detecting activation of this protein under various conditions. Until one day, I could not. All of a sudden, I could no longer see it! I would repeat my experiments over and over being extra careful with each step, but I was no longer able to see the shift in the protein. Talk about frustration and embarrassment!

Yes, embarassment. I always prided myself in my ability to do my experiments diligently, and when I couldn't, I felt like a failure. And because of this, for about two weeks, I kept my problem to myself. I was embarrassed to tell my labmates and especially my supervisor. But soon, my meeting with my supervisor was coming...so I had no choice.

Now, at this point, I should say that I feel blessed to have had such an amazing supervisor (Dr. Margaret Chou at the University of Pennsylvania). She was hard-core, but incredibly supportive, kind, and nurturing. Why did I feel embarrassed to share my problem with her? I guess I felt like I let her down.

Then came the big day when I revealed my failure to her. Her response immediately made me feel relieved. She shrugged it off as not a big deal, saying things like this happen all the time, and instead worked with me to help solve the problem. She first asked another member of the lab to try the experiments in parallel with me, and then we split the samples in half. I then ran mine and my labmate's samples side-by-side on my gel, while they ran our samples in parallel on their gel. The result...neither of us could detect activation from either sample.

This meant it was not a personal problem. I was not doing anything wrong, per se. And this was a huge relief to me. Margaret then asked me to compare all of the reagents used in the experiments and see if anything has been changed (or recently purchased). Starting at the beginning, I checked each one. Finally, I saw one. We recently purchased a new bottle of SDS (sodium dodecyl sulfate, which is a denaturing agent we use in our gels). And, I realized that this was 95% pure rather than the usual 99% pure one we usually use.

Could this be it? No way, I thought. Changing the purity of this one chemical from 99% to 95% was sufficient to affect how this protein migrated on the gel?

I borrowed 99% pure SDS from a neighboring lab and tested it side-by-side with our 95% pure SDS. And yes...when I ran my old samples in the gel made with the 99% pure SDS, I could easily detect the shift...but with the one made with 95% pure SDS, I could not.

Moral of this story: it is often that experiments don't work, but don't keep it to yourself. It's not your fault. Work with your colleagues and/or supervisor and systematically try to understand why. You may be surprised how supportive those around you really are!

When a project fails

Yes, I have another story for you here...again from my graduate student days. I was working on another protein called JNK and recently purchased a new antibody that I could use to detect its activation on a gel. As with any new reagent, I first tested with positive and negative controls, which seemed to suggest it was working fine. Then it was time to test it under the conditions I was investigating: how the environment outside of the cell -- the extracellular matrix -- may regulate this protein.

To my surprise, I seemed to show for the first time that these molecules were activating JNK! I was so happy! I was developing this project with many great results and then submitted it to an upcoming conference that I was looking forward to presenting at. I was ready to share my exciting results with the research community!

When testing different inhibitors in my experiments, I realized that an inhibitor of a protein called MEK seemed to block the activation of JNK. Hmmm...that is not right. MEK is upstream of a protein called ERK, not JNK. I then started to worry...was my antibody recognizing 'active JNK' actually detecting ERK?

I then tested this by isolating both JNK and ERK in parallel from the cells (immunoprecipitation using tried and true antibodies), and then tested my 'active JNK' antibody against both. To my dismay, the 'active JNK' antibody reacted with both. This means, this whole time I was likely looking at activated ERK, which had previously been shown to be activated in the conditions I was exploring. Nothing new.

I validated this with a few more experiments and confirmed that this project that I had been working on for a while now was a fallacy. It had failed.

What do you do when several months are wasted? When a project you were going to present as a poster at an upcoming conference has failed. Rest, recover, and recoup.

I took a week off to just rest and give myself time to recover. And then, working with my amazing supervisor, we came up with a new project for me to work on...one that led to my completed thesis and 3 published articles! In other words, it is perfectly fine to have a failed project. Learn from your experience as to why it failed to grow into a stronger and more resilient researcher going forward.

On a side note...you may be wondering...what happened with the poster presentation? I presented about the failed antibody with my many controls demonstrating its cross-reactivity with another protein to serve as a word of warning for other young researchers to be extra vigilant when using new/untested resources.

Moral of the story: when a project fails -- rest, recover, and recoup. It happens to everyone. Have 2 or 3 projects that you are working on in parallel so that if one fails, you have another to rely on. You may find that your 'backup' project becomes the most successful one!

When personal issues affect your research

We are not robots. We are humans with human experiences and feelings. And sometimes this can affect how well or efficiently we can do our research. That is fine. We should embrace our humanity and be practical in our professional lives. Let me share two more stories along these lines...

A Singaporean Journey

During my first post-doc in Singapore, I had an important family issue that needed to be taken care of back in the US. My post-doctoral advisor (Dr. Edward Manser at the Institute of Molecular and Cell Biology) was incredibly supportive and told me to take the time I needed...my position and my project would be waiting for me when I returned.

And my previous graduate supervisor (Margaret, from the above stories), was able to arrange for me to work for a short while in her lab while I settled this family issue in Philadelphia.

While the support from Ed and Margaret was absolutely amazing, this led to a gap in my research productivity as well as the emotional and mental strain that made it a real challenge to get things going again once I returned to Singapore. Because of this, you will see a big gap in my CV in my article output as I was dealing with these issues.

Did this affect subsequent interviews? Definitely. This often came us as to why there was a gap in my research output. Therefore, I started to explain in my cover letter what I had gone through to explain this gap, but also emphasized that after overcoming these issues, my productivity resumed (hoping to instill confidence in me as a researcher). Thankfully, being open about my personal issues was welcomed during my interviews and did not affect my job opportunities.

Moral of the story: when personal issues strike, take time to work through them and then be honest about them going forward. We are all humans and understand that we are not working in a vacuum. In being transparent, you may be surprised just how supportive the research community can be!

Memories of Japan

In my second post-doc, I was working for an amazing neuroscientist named Dr. Mineko Kengaku, first at the Brain Science Institute (RIKEN) in Tokyo, and then at Kyoto University. During this time, I was already married with hopes of having a child sometime in the future.

I had obtained a JSPS post-doctoral fellowship for 2 years and was working on an exciting project. Of course, it takes time to try a few projects to see which ones will pan out (I learned from my previous story!), but alas, I still managed to find one I really liked. As I was now in the last 6 months of my fellowship, Kengaku-sensei and I agreed on the project. She also managed to secure a position that I could apply for to remain at Kyoto University to continue on the project.

Around this time, my wife became pregnant and she wanted to return to Singapore to be around her family (as we had no family in Japan that could help us with our newborn). Understanding the importance of taking care of my new family, I then searched for research positions in Singapore that would allow us to move. I even flew there for a job talk at the National University of Singapore.

In this time, did I discuss this with Kengaku-sensei? No, I did not. I was worried that after she made the effort to secure a position for me, she would be upset. Instead, I hid it from her and did not tell her until the job offer from NUS came through. Needless to say, she was incredibly disappointed with me that I did not have discussed this with her first.

I also felt terrible as if I had betrayed someone who had been an amazing advisor to me.

We did move back to Singapore (and I then worked on my third and final post-doc)...but I did so under conditions that I really regretted.

With time, I apologized to Kengaku-sensei and realized that was very unprofessional and selfish. I should have been transparent and trusted her in the beginning, something I should have learned from the above experience.

Since then, my family and I have moved back to Japan (we love it here too much to stay away!), and I have made reparations with Kengaku-sensei and have visited her in Kyoto several times since...including earlier this year when I brought my daughter to meet her and my former colleague.

Moral of the story: learn from my mistakes and always be open and transparent with your colleagues (especially your supervisors) about personal issues that you are facing. Hiding them only caused frustration and regret for everyone involved.

Lessons learned

I hope that sharing some of my stories will help inspire and empower you to overcome any setbacks you experience in your research life. I think the most important common thread you will see is the importance of being open and transparent. You will likely be surprised how supportive your colleagues can actually be!