For most adults, the only opportunities we have to play with fluorescence is when we twist in a new light bulb or pick up a highlighter to mark an important section of a document.
But for Barbara St. Pierre Schneider, the Tony and Renee Marlon Angel Network Professor of Nursing at 51ԹϺ, and Alison Deng, a premedical biology honors student, fluorescing things is part of the regular research agenda. They stain white blood cells with fluorescent markers — the cellular type, not the Hi-Liter kind — and observe them under a confocal microscope to discover exactly where these cells travel in relation to damaged muscle fibers. They hope to someday uncover what the location and concentration of white blood cells in and around muscle fibers says about how the cells heal muscles.
The project kicked off in 2016, when St. Pierre Schneider became one of five co-recipients of a $30,000 51ԹϺ Faculty Opportunity Award (FOA) for a project led by primary investigator and life sciences professor Laurel Raftery titled “Quantitative Analysis of Cellular Interactions in Clinically Relevant Models,” which focuses on researching various cell-to-cell interactions.
Once the team received the funding in July 2016, Raftery notified her students of the opportunity to work on the research project alongside St. Pierre Schneider. Deng, who’d worked in biochemistry professor Bryan Spangelo’s lab the previous year, joined St. Pierre Schneider’s lab in the fall of 2016. Together, they’ve been blazing a trail toward discovery ever since.
St. Pierre Schneider: I’ve been interested in the white blood cell response to injury for a long time. The first patient I had as a nurse when I worked in the hospital was a woman with advanced cervical cancer. A colleague there explained to me how chemotherapy works. I was intrigued by how white blood cells responded to chemotherapy, and I started to develop an interest in the immune system. Fast forward to when I was a PhD student: A faculty member thought I could bring my interest in the immune system to muscle injury research, and that’s how I got started.
Deng: When Dr. Raftery sent out an email asking if anyone was interested in working on this project, I was interested because Dr. Raftery is a great teacher, so whatever she suggests, I’m curious about. I’d also taken a histology class that had to do with separately identifying muscle fibers and white blood cells, and I wanted to see it in practice, so I emailed back and said that I’d love to help.
Deng began her work in St. Pierre Schneider’s lab with the standard round of safety training, moving on shortly thereafter to work one-on-one on the fluorescent cell-staining technique with St. Pierre Schneider.
The staining effort wasn’t without its challenges. White blood cells actually auto-fluoresce when they’re active, as is the case when there’s inflammation in the body. Deng reviewed existing literature on staining with fluorescence, noting other researchers’ approaches and protocols. She experimented with cold fixing solutions and different mounting mediums to find would work best for her project.
But even after identifying an optimal staining technique, Deng still had to find the right tool to view her results. Since conventional microscopes can’t eliminate auto-fluorescence well, Deng would need to work with a confocal microscope to see the effects of her staining efforts. This powerful microscope increased optical resolution, narrowed the depth of field, and most importantly subtracted glare and noise from the background so that Deng could identify the structures she’d stained. Deng picked up the confocal microscopy piece from a user training session and some coaching from life sciences research associate Sophie Choe in the 51ԹϺ Confocal and Imaging Core.
Deng: I’ve learned a lot through this research so far. The simple fact that muscle fibers auto-fluoresce was surprising. I also learned that writing down everything is important—even what you read, like the literature and the complications that might come about in an experiment and how to overcome them.
St. Pierre Schneider: I commend Alison for her confidence and ability to master the techniques quickly. She’s now in a phase where she is critiquing her own work. It’s great to see that. That’s the life of a scientist: to take that initiative, to come in and be curious, and to be able to evaluate and improve upon what you do.
When Deng began her studies at 51ԹϺ, she had already decided to pursue premedical biology, with the goal of improving and saving people’s lives. But her time in the lab has opened her eyes to other ways she might go about accomplishing this same goal.
Working alongside St. Pierre Schneider, Deng sees the impact their research can make in the future. She’ll graduate in May but is taking a gap year to continue working in the lab before she goes on to pursue both MD and PhD degrees. It’s a good thing, too, since the work on this project has only just begun.
St. Pierre Schneider: We’re still in the early stage of this research. We have a long way to go. But because it’s early-stage, our project isn’t highly fundable yet. This is why seed money is so important, even though it’s challenging to acquire. Science is built upon formulating hypotheses based on existing data, but when you’re exploring new avenues, you may not have substantial data to formulate hypotheses with so you can pursue outside funding, so you have to do the preliminary work first. We were lucky that 51ԹϺ gave us the seed money to begin the project, but funds are running low now, and it’ll be a while before we have the preliminary data we need to formulate our hypothesis and pursue additional funding.
Deng: Right now, I get paid an hourly wage to do this research, but that would’ve ended if Dr. St. Pierre Schneider hadn’t figured out another way to keep compensating me. I would’ve volunteered if necessary, though, because what we’re researching is really cool, and I want to keep working on it.
St. Pierre Schneider: I know that many students volunteer, and that’s wonderful, but I think that, given the complexity of the work and what it means to the faculty member, it’s most appropriate to provide some compensation to students doing research.
Deng: I’ve learned a lot at 51ԹϺ about the importance of research funding. For example, in my phrenology class, we talked about how the research needed to create a certain vaccine wasn’t funded because the disease hadn’t become a threat to the U.S. yet, though it was in other parts of the world. But eventually, the disease reached us, and we didn’t have a vaccine, so it cost a lot of lives. If we don’t get funding for important things like research, there can be dangerous consequences.
For the time being, Deng will continue her research on white blood cells in muscle fibers, taking her work a step further by “stacking” her samples, using software to generate a 3-D reconstruction of muscle by placing multiple layers or sections next to each other to observe white blood cells along an entire length of muscle fiber.
Come summertime, St. Pierre Schneider will transition Deng to a U.S. Department of Defense-funded project, where Deng can apply what she’s learned from her current lab experience to study the effects of air transport on soldiers’ muscle injury and recovery.
St. Pierre Schneider: I’m really appreciative of the opportunity to work with Alison and look forward to continuing to work together. In many ways, Alison has directed her own project. Very early on, we talked about the types of experiments that would need to be done, and she organized them and figured out how to go from step one to reach the end goal. It’s great when a student has the confidence to be able to work independently and think critically about the project, because that’s how you really engage with research.
Deng: It’s funny because Dr. St. Pierre Schneider talks about my confidence, but I’m only confident because I know that I can always ask her anything, and she prepared me really well. It’s just clicked.
