The essence of Biological Research is asking questions that have never been answered, troubleshooting when things don’t work as expected, and making sense of what you see !

Harikrushnan Balasubramanian, our next pathbreaker, Staff Scientist at the Janelia Research Campus ( Howard Hughes Medical Institute), helps researchers design and carry out advanced microscopy experiments that can effectively answer their research questions.

Harikrushnan talks to Shyam Krishnamurthy from The Interview Portal about stumbling upon research groups working with fluorescence microscopy, and realising that the microscopy images were not only visually striking, colourful, and detailed, but also have immense potential as a powerful tool for scientific discovery.

For students, careers are not a race. People start at different points, get different opportunities, and move at different speeds — so comparing yourself to others is rarely useful or fair. The only meaningful comparison is with your past self.

Harikrushnan, what were your initial years like?

I was born in Chennai, but grew up across India in Chennai, Hyderabad, Mumbai, and Indore. Therefore, I got to experience many different cultures and cities which gave me a broader perspective on life. My dad worked in State Bank of India and my mom is a homemaker.

As a child, I was an avid reader. Science fiction novels and the “Young Discoverer” book series (Discovery Channel) opened my eyes to the wonders of the world, from the vastness of the cosmos to the tiny, invisible world of cells and microbes. I also enjoyed watching movies – Jurassic Park blew my mind with the thrilling idea that science could bring dinosaurs back to life. Cartoons like Pokémon sparked my imagination too. My parents further nurtured this curiosity by taking me to science museums and planetariums. Through all these experiences, I developed an aspiration to become a scientist.

I pursued the science stream in 11th and 12th standard. I’ll be honest – I struggled with mathematics, physics, and chemistry. They felt abstract and hard to connect with. But I liked biology. So, I decided to follow that path for my undergraduate studies.

What did you do for graduation/post graduation?

After 12th standard, I wanted to pursue a biology-related degree. In India, there’s a strong perception that the only meaningful career path for biology students is medicine. Since I wasn’t interested in that route, I explored alternative options and chose to pursue a degree in biotechnology.

Like every other student, I appeared for competitive entrance exams such as IIT-JEE and others – and flunked most of them! But I managed to get into SRM University in Chennai, and did my B. Tech in Biotechnology there. Happily, it turned out to be a great experience.

After finishing my undergrad, I knew I wanted to stay in science. I also knew that for a research career, a PhD is essential. So I decided to skip a master’s degree and go straight to a PhD (that would save me two years). Many countries allow you to apply for a PhD directly after a bachelor’s degree. At that time, such direct PhD opportunities in India were relatively limited, so I also explored options abroad in the US, Europe, and Singapore.

The GRE (and TOEFL) is required for admission in many foreign universities, while Indian universities require GATE. Some universities in Singapore and Germany also accept GATE. I appeared for these exams, and the strong scores I achieved helped support my applications. I attended a few interviews at Indian institutions like IISc and IIT-Madras but only got into one – the M. Tech Nanotechnology program at Anna University which I even enrolled into. However, I later received an offer for the Biological Sciences PhD program at the National University of Singapore (NUS), and I dropped out of the M. Tech program. No regrets there.

For students thinking about the PhD path: strong exam scores help get your application noticed, but they don’t guarantee admission. Your CGPA, prior research experience, and overall profile matter too – especially for international programs where evaluation is more holistic and not just based on academics. Reaching out to potential supervisors can also make a significant difference – a professor who already knows you and finds your profile interesting is far more likely to advocate for your application. Apply broadly, apply everywhere, and always have backups.

Coming back to my story, the NUS admission turned out to be a major turning point for my career. NUS is a top institution with a strong international research environment. Singapore is also geographically close to India, and is a safe and convenient city to live in. It felt like the right move and I moved to Singapore. I completed my PhD in an interdisciplinary lab focused on using fluorescence microscopy to answer biological questions.

What made you choose this career in Biotechnology?

One of my early inspirations was Dr. A.P.J. Abdul Kalam, the renowned scientist and former President of India. Seeing a scientist rise to such a prominent position was very motivating. His passion for science and his unwavering commitment to engaging with students, right until his death, left a deep impression on me. He embodied the idea that a scientist can be a source of hope and inspiration for an entire nation.

My interest in microscopy, however, developed in an unexpected way. During school and undergrad, microscopy was just another lab activity, like peering at cells or bacteria through a basic compound microscope. Nothing particularly special about it. Then, when I was searching for PhD labs to join, I stumbled upon research groups working with fluorescence microscopy. The microscopy images were visually striking – vivid, colourful, and detailed. Since I have always enjoyed art as a hobby, this immediately caught my attention. It felt like a unique combination of science and visual creativity. It wasn’t until I started my PhD that I realized how microscopy is probably the only tool where you can actually see biology in action at the smallest scales. That’s when I began to truly appreciate its immense potential as a powerful tool for scientific discovery.

Sometimes you find your career in the most unexpected ways. Keeping an open mind and exploring different possibilities can lead you to paths you may not have initially considered.

How did you plan the steps to get into the career you wanted? Tell us about your career path.

I completed my PhD in biophysics, specializing in fluorescence microscopy. My research focused on understanding the epidermal growth factor receptor (EGFR), a protein on the surface of cells that plays a critical role in how cells communicate, grow, and divide. When EGFR malfunctions, it can drive the development of cancers, which makes understanding its behaviour medically important. The core question my research addressed was: how does EGFR organize and interact on the cell membrane? While this may sound abstract, it has very real consequences. Whether EGFR molecules act individually or cluster together, how dynamically they move within the membrane, and what hinders or promotes that movement – all of these factors can strongly influence the strength and outcome of cellular signalling. When these processes are disrupted, signalling can become abnormal, and that is one of the ways diseases like cancer develop.

Answering these questions required an interdisciplinary approach, combining biology with physics, chemistry, and mathematics – ironically, the very subjects I struggled with in school! However, I found that applying them in a practical biological context made them much more intuitive and meaningful. You don’t need to be a textbook expert in these subjects; you just need to understand enough to apply them effectively. When you see how a concept solves a real problem, it stops being abstract and becomes useful.

Toward the end of my PhD, I realized that my interests had evolved. I had lost the drive to do my own independent research and didn’t want to become a professor (which, again, is what a lot of people assume is the only career path after a PhD). I still wanted to do science, just in a different way. I had microscopy skills, and enjoyed teaching and helping other scientists use it effectively. So, I looked for career paths where I could do exactly that – apply microscopy across many different projects and support researchers in doing so.

That search led me to an incredible opportunity in the US at the Advanced Imaging Center of Howard Hughes Medical Institute Janelia Research Campus. They offered a postdoctoral-level fellowship specifically designed to train scientists for careers in core facility roles, where you as a microscopy expert support other researchers with their imaging experiments. It felt like a perfect fit.

I applied, got in, and made the move to the US. Since completing that fellowship, I have continued working in the same institution as a full-time staff, focusing on enabling and supporting scientific research through imaging technologies.

How did you get your first break?

I would say I had two major “first breaks” in my career.

The first was getting into the PhD program at NUS. I had applied to several programs across India, US, Europe, and Singapore, but only NUS came through with an offer. That break gave me access to a strong research environment and helped me build the skills I needed for a career in science.

The second was getting selected for the fellowship at HHMI Janelia. I just stumbled across it while searching for opportunities that matched my skills and interests. The timing was right, my microscopy background was a good fit, and fortunately my application was successful. That fellowship ultimately led me to my current role.

In both cases, a lot of it was being in the right place at the right time, with the right qualifications. Luck matters but also favors the prepared – you can’t always control when a door opens, but you can make sure you’re ready to walk through it when it does. And I think most people who’ve had a good break in their career will tell you the same thing.

What were some of the challenges you face? How did you address them?

There were many challenges along the way. Here are a few key ones:

Challenge 1: Choosing a non-traditional career path

In India, biology is often closely associated with medicine, and choosing a different path with an uncertain future can be difficult. There is social pressure, unsolicited opinions, and a lot of “but what will you do with that degree?”

I evaluated my options and chose the path that aligned best with my interests. Having supportive parents also made it easier to commit to this decision.

Challenge 2: Moving abroad

Leaving India to start my PhD in Singapore was exciting but also challenging. I’m an introvert and socially shy. Adjusting to a new country, culture, and academic environment, without family support nearby, was not easy. And this was coming from someone who had already lived across multiple cities in India, which is more than what many people experience. However, I had never lived independently in India, so managing everything on my own, from cooking and chores to grocery shopping, was a major transition.

I gradually built a support system through peers and mentors at NUS, made friends from all over the world, and learned to be independent. Discomfort is temporary, but growth is permanent and comes from stepping outside your comfort zone.

Challenge 3: PhD struggles

Nobody truly realizes how hard a PhD is until they’re doing it themselves, even if they’ve seen others go through it. There were periods where experiments didn’t work and progress felt slow, which led to self-doubt of whether I was cut out for this. This is often referred to as imposter syndrome – the feeling that you’re a fraud who is not good enough and doesn’t belong, that everyone else knows more than you, and that you somehow fooled everyone into letting you in.

Almost every PhD student experiences this. Knowing that doesn’t necessarily make it easier – you still feel terrible, isolated, and discouraged. What helped was my lab friends and colleagues. We had daily tea breaks where we’d rant about our lives, share setbacks and frustrations, laugh about it, and then go back to the lab feeling re-energized and ready to fight another day. When you hear that everyone has the same problems, even if nobody has a solution, the struggle feels more manageable. In the end, a PhD teaches resilience more than anything else – success is less about raw talent and more about perseverance.

Challenge 4: Redefining my career direction after PhD

Toward the end of my PhD, I realized I did not want to pursue the traditional academic path of becoming a professor, which created uncertainty about what to do next. Alternative career paths are simply not well-advertised or discussed in academic circles. Only when you actively explore outside that bubble do you realize that other paths exist – probably not many, probably not easy to get into, but they are there.

Where do you work now?

I currently work at the HHMI Janelia Research Campus – one of the world’s top institutions for biological research and advanced microscopy – in the United States. I am a staff scientist in their imaging core facility that supports researchers working on a wide range of biological questions. Our facility hosts 15+ microscopes and serves ~150 users.

What problems do you solve?

I help researchers design and carry out microscopy experiments. They come to me when they need to see biology happening at the microscopic scale – things like a protein moving inside a cell, a neuron firing in a brain, a microbe infecting a cell, and so on. I help them figure out how to image their samples by choosing the right microscope, setting up and optimizing their experiments, troubleshooting when things don’t work as expected, and making sense of what they see. My job is to make sure the microscopes are working properly and that every scientist, regardless of their experience level, can use them effectively to answer their research questions.

What are the skills are needed for your role? How did you acquire the skills?

The job requires a mix of technical and people (not the introvert’s natural habitat) skills. On the technical side, you need a solid understanding of microscopy, biology, and data analysis – all of which I built through my academic training. Equally important are communication skills, patience, and the ability to explain complex concepts in a simple and practical way to those outside the field. These “soft” skills are often underemphasized during a PhD, which tends to prioritize technical training. My fellowship training and the actual experience of doing the job helped me develop them.

What’s a typical day like?

A typical day involves doing microscope maintenance and performance checks, project consultations with researchers, training users with varying experience levels, troubleshooting experiments, and helping with data interpretation. But there is no fixed routine. One moment I might be fixing a technical issue on a microscope, and the next I’m sitting with a user helping them understand why their images aren’t looking right.

What is it you love about this job?

I love that I get to use the skills I’ve spent years developing and honing, rather than setting them aside as many people often have to when they change roles. I enjoy the variety of contributing to many different scientific projects at once, instead of being confined to a single narrow research question.

It is also deeply satisfying when researchers are able to successfully use our microscopes and the imaging contributes to new scientific discoveries. Many users are new to microscopy, so we typically sit down together and try to figure out how to answer their scientific questions. This helps them see what the microscope can actually do and how to get the most out of it, rather than leaving frustrated. Users appreciate simple, practical guidance that helps their biology make sense, not overwhelming them with technical details of the microscope. It’s a two-way street: I learn something new from every user, and they pick up something useful from me.

Beyond the science, I value the overall work-life balance this role offers. While the pay is lower than in industry, it comes with a regular 9-5 schedule, weekdays only, strong job security, low stress, and generous vacation time. A fulfilling career isn’t just about the work; it’s also about having a life outside of it. You work to live, not live to work.

How does your work benefit society?

My work supports scientific research by helping researchers use advanced microscopy to better understand how biological systems function. This includes studying processes such as how cells behave, how the brain works, or how diseases develop – knowledge that ultimately contributes to advances in medicine and biology. Science is a team sport, and core facility staff are an essential part of that team, even if we’re rarely in the spotlight.

Working at Janelia gives me a unique vantage point to see how rapidly microscopy is evolving and how new technologies are expanding what scientists can observe and measure. An important development is the growing role of artificial intelligence (AI) in microscopy. AI can help design experiments, automate data collection and analysis, and extract deeper insights from complex datasets. When used carefully and with high-quality data, it has the potential to significantly accelerate scientific discovery that can lead to future breakthroughs.

I also extend the impact of my work through teaching and training (like in the photo below). This happens at multiple levels, from one-on-one sessions with individual researchers at the microscope, to organizing and teaching at larger microscopy workshops that bring together scientists from across institutions and even different countries. When I teach, I am not simply helping to solve a single experiment; I am building long-term capability. This creates a ripple effect: individuals I train go on to advance their own research and often pass that knowledge on to others. In this way, teaching quietly becomes one of the most powerful ways scientists can contribute to society.

Tell us an example of a specific memorable work you did that is very close to you!

One piece of work that I’m particularly proud of is a research paper I co-authored during my PhD. Let me explain what we did in simple terms. When scientists study living cells, they often want to understand multiple things: What does the inside of a cell look like in fine detail? How fast are the molecules moving? How many are clumped together? What else are they doing? The challenge is that different microscopy techniques are usually needed to answer each of these questions. Each method is optimized for one type of information, so getting the whole picture requires running multiple experiments, collecting separate datasets, and spending a lot of time analyzing them. Crucially, there is no reliable way to resolve discrepancies when the results from different techniques don’t quite agree with each other.

In our work, we showed that it is possible to extract a lot of information (structure, movement, and molecular organization) from a single imaging dataset. In other words, one experiment could give multiple answers at once. This is done by analyzing the same dataset in different ways. Each analysis focuses on a different aspect of the data, allowing us to pull out different kinds of information from the same experiment. And because everything comes from the same data, you can use one type of result to cross-check and improve another. For instance, we used information about how molecules were moving to identify and remove false structures (features that looked real but were actually artefacts) that appeared in high-resolution images. This made the overall results significantly more reliable and trustworthy.

The work took over 1.5 years and involved an interdisciplinary team – microscopy experts, cell biologists, and computational scientists – all working together. Complex problems are often best solved by combining different perspectives and skill sets. Seeing everything come together and eventually get published was very rewarding. Knowing that the work has since been used and cited by researchers around the world makes it one of the most satisfying moments of my scientific career.

Your advice to students based on your experience?

Students already get a lot of advice (often unsolicited) from left, right, and centre. So let me pitch in too with my two cents.

  1. Don’t blindly chase “safe” careers: explore first

In India, everyone talks about the “safe” options: medicine, IT, government jobs. But they’re not easy to get; crores of people are chasing the same opportunities. And even if you do get it, you may realize it doesn’t suit you. So explore a bit first. Sometimes the better path is the one fewer people are looking at, where you can actually be good and stand out.

  • Marks matter: but only for the next step

Marks matter… briefly. They will help you get through the next door. After that, nobody cares. What matters far more is whether you can solve problems, learn new things quickly, communicate clearly, and work well with others. So do well enough to grab the next opportunity, but don’t let your marks define your identity or self-worth.

  • Your career will probably not go as planned: and that’s okay

Very few careers follow a straight line. Mine didn’t. Sometimes you choose a path deliberately; sometimes you stumble onto one and find you like it. Both are valid.

The world is also changing faster than ever (AI is already accelerating this): some jobs will disappear and new ones will emerge. So don’t get fixated on a specific job that might not exist tomorrow. Focus on learning, adapting, and building transferable skills. And remember, opportunities don’t come just because you’re talented. People need to know you exist. Network widely – talk to people, build genuine relationships, and put yourself in the right environments. Who you know is equally, if not more, important than what you know for career progression.

  • Don’t compare your timeline to others: everyone’s journey unfolds differently

Careers are not a race. People start at different points, get different opportunities, and move at different speeds — so comparing yourself to others is rarely useful or fair. The only meaningful comparison is with your past self. If you have grown, learned, and made progress relative to who you were, you are doing well.

That said, it’s easy to lose sight of this when people around you seem to be moving ahead in life faster. Self-doubt creeps in, hard periods pile up, and if left unaddressed, it takes a toll on your mental health. So step back, seek support, give yourself time to reset, and fix what you can and let go of what you can’t. Your mental health matters enormously. Take it seriously and protect it actively.

  • Think beyond the job: environment and balance matter

A job is not just the work itself. Your manager, team, and work environment shape your daily experience. A good environment can make even challenging work enjoyable. A bad one will make even an exciting job feel exhausting and demoralizing.

There’s a popular Japanese concept called ikigai (“a reason for being”): the idea that your job, passion, and life purpose should all align perfectly. Sounds wonderful and ideal. But not everyone is lucky enough to find that dream job. Sometimes your ikigai is just a combination of different things that work for you, not one magical job. You can have a stable job that pays well and pursue your other interests outside of work. What matters is balance and long-term sustainability – don’t burn yourself out chasing something unrealistic.

Future Plans?

I plan to continue growing in my current direction: building a long-term career managing shared research infrastructure and helping scientists get the most out of cutting-edge imaging technologies. I also hope to improve access to these tools for everyone, through training programs, workshops, and collaborations. Science should not be a privilege of well-funded institutions alone.

I’ll also take my own advice here. I’ve told you not to get fixated on a specific plan, because the world changes and so should you. I’ve said to keep learning and adapting, to stay open to unexpected new paths, and to not sacrifice balance for unrealistic ideals. I intend to follow all of that myself. Whatever comes next, I want to make sure I’m growing relative to who I was yesterday. The rest, I’ll figure out as I go.

And here’s a parting gift: a fluorescence microscopy image of a cell, with actin – the cell’s internal “skeleton” – lit up in colour. Pretty cool, isn’t it? It’s as much art as it is biology, and a small glimpse into the microscopic world that scientists like me get to explore every day.

You can also read a previous, more specific interview on my current career here:
https://focalplane.biologists.com/2024/10/02/an-interview-with-dr-harikrushnan-balasubramanian/