Powerful microscopes advancing research for students, faculty

Ryan Gabriel, '07, (l.) and Amy Hart, '08, use the confocal microscope for their research. Most confocals are located in medical schools or large research universities.

Whether studying cancer cells, Chesapeake Bay sponges or lymphatic hearts in amphibians, campus researchers are saving time and getting better images from three powerful University microscopes.

Located in the Biological Imaging Suite of Gottwald Center for the Sciences, the microscopes are used by classes and individual researchers, said Carolyn Marks, director of biological imaging.

With the most resolving power of the three, the JEOL 1010 transmission electron microscope (TEM), shines an electron beam through a sample, revealing cross-sections of bacteria and other organisms. Recently upgraded with a bottom-mount AMT camera system, it can capture images magnified up to 300,000 times.

The JEOL 6360 scanning electron microscope (SEM) looks at surfaces. It was used last summer in the course Unseen Life and has been used by students in Val Kish’s Biology of Cancer course. The SEM also has an X-ray analysis system that allows the user to “see” and map out the elemental composition of samples.

A Leica SP2 laser-scanning confocal microscope (confocal) is usually found only at medical schools and large research universities, “but even then students have to wait for training and wait to use the instrument,” says Marks. The confocal works with lasers and a fluorescence technique called epifluorescence that allows it to see “slices” of a sample.

Marks says that the imaging suite also contains all needed ancillary equipment to prepare a wide variety of samples for all three imaging modalities. “If the faculty supply the reagents and consumables, I will personally train their students on the proper use of the prep equipment and the microscopes in the lab.”

Amy Hart, ’08, of Rumson, N.J., used the confocal last summer to study circadian rhythms of the fly brain. She says “It’s great the University lets us use the confocal—other universities have just the higher-ups using it.” Hart, who spent 10 weeks conducting research under the guidance of John Warrick, assistant professor of biology, looks forward to a career as a researcher.

Another student, Gerard Gomes, ’08, of Houston, Texas, has been using the confocal “to observe the fluorescence label corals I prepare in my lab. An enzyme called urease is marked with a florescent label to see where it is located in the coral.” Gomes works out of professor Roni Kingsley’s lab.

Ryan Gabriel, ’07, of Ventnor, N.J., who works in professor Val Kish’s lab, is studying brain cancer metastasis and uses the SEM to magnify cells up to 11,000 times. He also uses the confocal to study human glioma cells with a “stain that will show a picture of its actin cytoskeleton.” The confocal, he says, “is very useful because it provides a visualization into the cell.”

Nicole Buell, ’06, of Raleigh, N.C., says she used the TEM to examine the effects of global warming on Chesapeake Bay sponges’ bacterial communities. Working last year with Nathan Lemoine, ’06, of Houston, Texas, and Malcolm Hill, associate professor of biology, she took pictures of bacteria at 5,000–20,000 magnification to see changes in tissue possibly caused by the stress of exposing it to higher temperatures. 

Over the summer, she examined “the transmission of bacterial symbionts into larvae from adult sponges.” She used the TEM to examine adult tissue with larvae attached as well as to get shots of single larvae.

Faculty also use the microscopes. Gary Radice, associate professor of biology, has been using the confocal to study “the development of tiny hearts that pump lymphatic fluid in amphibians, reptiles and birds.” He says he is trying to find out “where and when during embryonic development these lymphatic heart muscle cells form and what signals the embryo uses to tell the cells when and where to grow.”

The confocal, he says, “provides greater resolution with thick specimens, such as a whole frog embryo, and allows one to make images in 3D. With older techniques, I would have to cut the embryo into a dozen or even hundreds of sections, photograph each section individually and then painstakingly reconstruct the 3D view. It might take several weeks. The confocal does the same thing more accurately in a few minutes.”

Craig Kinsley, professor of neuroscience, has used the confocal for “precise examination of fluorescent-stained tissues, to examine triple-labeled neurons. This project will ultimately allow us to understand, for instance, what happens in the brain of a female rat to convert her into a mother.”

Kinsley also uses two Carl Zeiss microscopes, an Axioplan and an Axioskop, in his lab. “Both are upright, light microscopes” that he uses to do image analysis and computerized microscopy.

“These microscopes are absolutely necessary to the work that we do,” he says. “They couple the behavior that an animal displays to its regulation by the brain. In the process, my students get a chance to glimpse the awesome complexity of—and to understand an infinitesimally small part of—the brain and nervous system, our personal universe.”