Of the five seniors on their team, only Micah Hagedorn says he thought they had a shot at the Best in College award — the top honor at the College of Engineering and Computer Science’s annual Senior Design Competition — and that was only after the team earned a nod for the best project from the Mechanical Engineering department. Just weeks earlier, things were not going well for Hagedorn and teammates Nicole Kormos, Rosa Carapia, Kenny Conuel Oralde and Emmet Reamer. Multiple times they’d had shipments of biological materials spoil when the supplier mistakenly shipped them to the Ann Arbor campus. And Carapia spent weeks trying to figure out their not-so-state-of-the-art microscope — at one point resorting to contacting the rep whose business card had been attached to the device who knows when. “It was the last couple weeks and I was, like, ‘Oh my gosh, this isn’t going to happen,’” Carapia says. “I was really thinking, ‘Our presentation was just going to look dumb because there’d be nothing there.’”
The team bumped into quite a few challenges, in part, because their multi-faceted project was one of the more ambitious in the competition. Assistant Professor of Mechanical Engineering Caymen Novak had it on her to-do list for some time to bring an imaging technique known as traction force microscopy to the Dearborn campus for the first time. TFM is used often in mechanobiology to study how cells interact with their microenvironments, and Novak thought it could be very useful for her current work, which is investigating how sex-based differences influence pulmonary fibrosis, a lung disease marked by significant scarring and stiffening of lung tissue. “So just to explain it briefly, you have a gel with fluorescent beads in it, and you put cells on it, so the cell interacts with the surface and pulls on it,” Novak explains. “Then, you take some ‘before’ pictures of the cells and the fluorescent beads, then you lift the cells off and take an ‘after’ picture. By measuring the movement of the beads, you can get a representation of the amount of force the cell is exerting on the surface.”
Novak had used this technique in her postdoctoral work at The Ohio State University, but there, she was plugging into an established lab setup. She hadn’t ever personally created the gels or configured the microscope for this type of imaging, and the analysis protocol was a closely guarded secret of the project’s principal investigator. So when Kormos, who’d been working as a student researcher in Novak’s lab, asked Novak if she had any projects for her and her Senior Design teammates, Novak immediately thought of the TFM setup. “I thought, ‘This sounds like a really ambitious Senior Design project. Let’s see how far they get,’” Novak says. Kormos took the idea to her teammates, who all liked the idea. They sketched out a plan for who would do what and got to work.
Because TFM is an established technique, there was actually quite a bit of literature out there to guide them. But it’s hardly a plug-and-play technology. The gels, for example, can’t be purchased off the shelf. You have to buy all the ingredients and make your own gel from scratch, fine tuning the chemistry so you have a medium with the proper stiffness for the kind of cells you want to study. Kormos and Reamer took on that part of the project and ran into several challenges. “You’d think because this has been done before, it would be pretty straightforward, but you follow the recipe, and sometimes your gel just doesn’t form,” Kormos says. “So we had to do some digging and figure out which component was doing what. Then we learned you had to add this component before that one or it wouldn’t work, or you have to dilute something just before you add it. So it took some troubleshooting before we found the proper protocol.” And then there was the unexpected challenge of even getting the materials properly delivered to their lab. Despite specifying the correct Dearborn campus address, Reamer says the distributor shipped their biologically sensitive components — one costing $400 for 50 milligrams — to the Ann Arbor campus not once but twice. When the third shipment finally made it to the lab, it arrived a week late. “I spent a lot of time on customer service,” Reamer says, wryly. “That was probably my biggest contribution to the project.”
Carapia, meanwhile, was wrestling with the lab’s less-than-ideal microscope to see if they could get it to work for TFM. She got some initial guidance from a couple other researchers on campus who also use this particular instrument. She made some initial progress — only to discover that she’d need to integrate a totally different camera-software setup than the one she’d just spent the past few weeks learning. Then, a weeks-long email back-and-forth with the person on that business card ended up in a dead end. In the end, Carapia relied on her engineer’s instincts, rolled up her sleeves and figured out most of it herself.
Hagedorn and Oralde tackled the analysis part of the project. Essentially they would have to write and tweak software to properly measure the displacement of the fluorescent beads and then convert those measurements into forces, given the known characteristics of the gel. Hagedorn dug into the published literature and found an open-source algorithm he thought they could work with. “By the end, it was pretty good, but initially, we got a lot of random arrows that were pointing in random directions,” Oralde says. “And we had to tweak variables and figure out what the right contrast was for the images, so the algorithm was tracking points that were relevant and not just random,” Hagedorn adds.
All the effort finally — and somewhat unexpectedly — paid off. With just a week or so to go until the Senior Design Competition day — and following a 19-hour session in the lab — they got their final set of images to work, measured the displacements and calculated the corresponding forces. The students say they would have loved to have had more time to run a mini-study with their technique, which was their original plan. (They joke it may have been possible had their FedEx packages arrived on time.) But they’re ultimately satisfied with the results. Novak is now digging through their final report to see what her next moves will be. “I’ve still not gotten hands-on with this myself, so I’ll have to see if I can make this process work, or possibly throw it to another Senior Design team to keep working on it,” Novak says.
Regardless, she’s impressed with the team’s hard work and tenacity. “It was interesting to watch them experience the difficulties of research,” Novak says. “They were, like, ‘We were there for hours trying to take these images.’ And I’m, like, ‘Yep, that’s how it works.’ But you have to admire their dedication in forcing this project to work on any level. In research, everything takes three times as long as you predict, often because of silly things, like deliveries going to the wrong address, which are totally beyond your control. And then you have to put way more effort in than you think. So that was a little eye-opening for them. But I’m sure they’ll feel it was worth it because they won everything! It doesn’t get better than that.”
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Story by Lou Blouin. Photos by Annie Barker.
