IU Gets New 3-D Microscope

A new laser-equipped microscope at Indiana University, Bloomington’s Light Microscopy Imaging Center makes it possible to examine biological samples with unprecedented detail in three dimensions.

The $1.2 million DeltaVision OMX superresolution microscope from Applied Precision LLC of Issaquah, Wash., was paid for entirely with funds from the American Recovery and Reinvestment Act of 2009, through a National Institutes of Health program that supports high-end instrumentation at America’s most deserving centers of higher education.

A fixed PTK (marsupial kidney cell line) cell in mitosis is shown. Condensed chromosomes stained with Hoechst stain are shown in blue, while microtubules labeled with an antibody to tubulin are shown in red. Imaging was done in the structured-illumination mode. More than 2000 images were taken and processed to create this image. (Image: Eric Workman and Jim Powers)

“It’s a fantastic and unique acquisition for our university,” said Claire Walczak, the Imaging Center’s executive director. “This superresolution microscope, one of only 16 in the world and one of only eight commercial units, is part of our vision to bring state-of-the-art technology to IU’s life science researchers, to enable them to address questions that they did not have the ability to ask previously, due to the lack of appropriate technologies.”

Walczak is a professor of biochemistry and molecular biology in the Medical Sciences Program in Bloomington, an arm of the IU School of Medicine. Walczak also holds an adjunct appointment in the IU biology department and is part of the biochemistry program.

The instrument is exceptionally fast in collecting images of a biological specimen, and this speed enables scientists to gather crucial data. The device uses laser light of four different colors to illuminate samples, while four extremely sensitive digital cameras capture images every 10 ms at the imager’s speediest setting. The device can produce as many as 5000 full-color images per minute for its major task of producing highresolution images. Known as a “structured-illumination” microscope, the device will help IU scientists attain a better understanding of how proteins are distributed inside cells.

Most high-technology light microscopes reach the limits of resolution at 250 to 300 nm — the diameter of a small bacterial cell. The new OMX microscope can produce clear images down to 100 nm in the lateral dimension. Resolution along the Z-axis (perpendicular, or coming out of the page) is somewhat lower but still tremendously improved relative to previous technologies.

“We’d envisioned this device would be most useful for microbiologists, cell biologists and neurobiologists at IU,” Walczak said. “But we expect scientists from many other fields will come up with creative ways to take advantage of it.”

Light Microscopy Imaging Center (LMIC) manager Jim Powers is responsible for training university researchers — as well as visitors — to use the device.

“The imaging center is a user-oriented resource,” Powers said. “Scientists rent time on our devices, and receive training to use them, but after that, we expect they’ll be able to work independently.”

Scientists on campus get a reduced rate when using the LMIC’s many microscopes, because of support from OVPR, the College of Arts and Sciences, Medical Sciences, and Optometry. At present, the OMX is still in a training mode in which Powers is working closely with Sid Shaw, an assistant professor of biology and the technical director of the LMIC, as well as university research staff to calibrate the device and establish protocols for future, similar uses. The LMIC staff expects the instrument to be available to all researchers on the campus shortly.

The arrival of the DeltaVision OMX microscope has spurred Walczak, Shaw and Powers to consider LMIC’s future needs. Partly because of the DeltaVision OMX’s size, the LMIC is now out of physical space. In addition, the device produces so much data (4000 images takes up about 1.5 GB of hard drive space), Walczak and Powers said one of the center’s next priorities is to improve the center’s information technology infrastructure through continued collaboration with the university’s Information Technology Services. Walczak and Powers want to ensure that the large data sets produced by the OMX imager can be stored rapidly — as well as protected from power outages and other catastrophes.

“We have some new things to think about, and lots of new things to see,” Walczak said.

For more information, visit: www.indiana.edu/~lmic