Confocal microscope acquisition opens new possibilities at IU Light Microscopy Imaging Center
Down the stairs near the first floor entrance of Myers Hall on the IU Bloomington campus, IU scientists are peering beyond what’s visible to the naked eye to reveal the complex hidden mechanisms that drive biology, chemistry and other processes in the natural world.
The IU Light Microscopy Imaging Center is home to nearly a dozen extremely powerful instruments capable of, among other things, tracing the delicate contours of the brain, mapping a chemical’s path though cellular channels or capturing the smallest details of an iridescent insect’s wing.
A 3-D image of a cladoceran, or water flea, created using the new instrument. Image courtesy Eduardo Zattara
Recently, the center acquired the latest in its collection of ultra-advanced imaging technology: a Leica TCS SP8 spectral confocal microscope system.
A confocal microscope uses a combination of intense laser illumination and ultra-sensitive light detectors to capture the interior details of microscopically small objects layer by layer, after which the images are automatically reconstructed to create a 3-D picture.
The high-powered instrument, which was acquired with support from the IU Bloomington College of Arts and Sciences and Office of the Vice President for Research, is the first upgrade of this particular technology at the Light Microscopy Imaging Center since 2007 – a significant leap in power and capability, including the ability to capture near-real-time images.
“Up until recently, the majority of imaging technology has only been able to provide scientists with structural information; the real cutting-edge is real-time imaging,” said Jeff Zaleski, a professor in the College’s Department of Chemistry and associate vice provost for sciences at IU Bloomington. “Today, our researchers need the ability to see changes in structural events in real time and watch processes as they happen — to view not just biological structures but also the actual physical processes that occur during those events. This new instrument moves us significantly further in that direction.”
The brand name of the new instrument may be familiar to photography enthusiasts, but the Leica TCS SP8 isn’t merely a high-powered camera – or even a typical confocal microscope. The instrument boasts myriad detectors and electronics not found in “off-the-shelf” systems.
A cutaway reconstruction of a growing beetle horn. Image courtesy Eduardo Zattara
Instead, IU’s custom-built system was designed to accommodate the advanced capabilities required by investigators at a major research university, said Sid Shaw, associate professor in the College of Arts and Sciences’ Department of Biology and technical director at the imaging center, who expects the new technology to provide services to IU departments beyond biology, such as chemistry and biochemistry.
“These commercial systems are sort of like cars; there is a base model that you configure for your particular needs,” Shaw added. “In our case, it’s more Ferrari, less Yugo.”
Since the instrument can capture near-real-time images, the Leica TCS SP8 is also better suited to research that uses live samples, said Jim Powers, an assistant research scientist in the Department of Biology, who serves as imaging center manager.
He also highlighted the instrument’s white light laser, which makes the system compatible with any fluorescent probe — the technology used to differentiate biological structures under investigation (a specific protein, for example) from the surrounding matter by causing them to light up, or “excite,” under laser light.
This feature is significant for the center because its earlier confocal microscope was limited to only seven probes. If a researcher came to the center with a painstakingly prepared sample — but neglected to use one of the correct probes — they simply could not image it. “Now, it’s pretty much unlimited,” Powers said.
LMIC Manager Jim Powers uses the Leica TCS SP8 spectral confocal microscope system. Image by Kevin Fryling
Among the first images captured using the new instrument are two beautiful, highly detailed pictures of insect structures created by Eduardo Zattara, a postdoctoral research fellow in the lab of Armin Moczek, a professor in the Department of Biology whose work concerns the evolution and development of beetle horns.
One image shows a cutaway reconstruction of a growing horn, a test image for an experiment investigating specific aspects of the horn’s development. The other shows a cladoceran, or water flea, collected from the lily pond by the Herman B Wells Library. The natural “auto-fluorescent” properties of the arthropods’ shell provides dramatic contours to the creature’s image as it rotates in 360 degrees.
It’s these sorts of capabilities that helped the Leica TCS SP8 win out over other similar instruments, four of which were physically transported to campus over the summer for interested researchers to “test run” the new technology.
The current system was officially installed at the Light Microscopy Imaging Center near the end of the fall semester. After a long wait, Powers is pleased to finally offer training sessions on the instrument. These personal sessions, which began recently, will extend through this semester and beyond.
Researchers interesting in training on the Leica SP8 can set up an appointment by emailing japowers@indiana.edu.
Video by Eduardo Zattara
