A new method of microscopy at UT Southwestern provides an insight into the future of cell biology: Newsroom

DALLAS – June 28, 2021 – What if a microscope allowed us to explore the 3D microcosm of blood vessels, nerves, and cancer cells instantly in virtual reality? What if I could provide views from multiple directions in real time without physically moving the specimen and running up to 100 times faster than current technology?

Red Fiolka, Ph.D. and Kevin Dean, Ph.D., are adjunct professors of cell biology and the Lyda Hill Department of Bioinformatics

UT Southwestern scientists collaborated with colleagues in England and Australia to build and test a new optical device that converts commonly used microscopes into multiangle projection imaging systems. The invention, described in today’s article Methods of nature, according to researchers, it could open new avenues in advanced microscopy.

“It’s a completely new technology, although the theoretical foundations of it can be found in the old computer literature,” says the corresponding author, Reto Fiolka, Ph.D. Both he and co-author Kevin Dean, Ph.D., are assistant professors of cell biology and the Lyda Hill Department of Bioinformatics at UT Southwestern.

“It’s like taking the biological specimen by the hand, turning it over and inspecting it, which is an incredibly intuitive way to interact with a sample. Quickly imagining the sample from two different perspectives, we can visualize the sample interactively in virtual reality on the fly, ”says Dean, director of the UTSW Microscopy Innovation Laboratory, which collaborates with researchers across campus to develop custom instruments that take advantage of advances in light microscopy.

Currently, the acquisition of 3D image information from a microscope requires a data-intensive process, in which hundreds of 2D images of the specimen are put together into a so-called image stack. The researchers explain that, in order to visualize the data, the stack of images is loaded into a graphical software program that performs calculations to form two-dimensional projections from different viewing perspectives on the computer screen.

“These two steps are time consuming and may require a very powerful and expensive computer to interact with the data,” Fiolka says.

The team realized that it could form projections from multiple angles by optical means, obviating the need to acquire stacks of images and render them with a computer. This is achieved with a simple and cost-effective unit consisting of two rotating mirrors that are inserted in front of the microscope system camera.

“As a result, we can do all this in real time, without any noticeable delay. Surprisingly, we can look at our samples from different angles “live” without rotating the samples or the microscope, “says Fiolka.” We believe this invention can represent a new paradigm for acquiring 3D information using a fluorescence microscope. “

It also promises incredibly fast images. While a whole stack of 3D images may require hundreds of camera frames, the new method only requires one camera exposure.

Initially, the researchers developed the system with two common light-sheet microscopes that require a post-processing step to make sense of the data. This step is called distorting and basically means rearranging the individual images to remove some distortion from the stack of 3D images. Scientists originally tried to make this deviation optically.

While experimenting with the optical deviation method, they realized that when they used the wrong amount of “deviation,” the projected image seemed to rotate.

“That was the aha! moment. We realized that this could be greater than a simple optical deflection method; that the system could work for other types of microscopes as well, ”Fiolka said.

“This study confirms that the concept is more general,” Dean says. “We have now applied it to several microscopes, including confocal microscopy of light sheets and a rotating disk.”

Using the new microscope method, they imagined calcium ions carrying signals between nerve cells in a culture dish and examined the vasculature of a zebrafish embryo. They also quickly imagined moving cancer cells and a beating zebrafish heart.

USSW co-authors include Bo-Jui Chang, Etai Sapoznik, Theresa Pohlkamp, ​​Tamara S. Terrones, Erik S. Welf, Vasanth S. Murali and Philippe Roudot.

Researchers at the MRC Molecular Biology Laboratory, Cambridge, UK; Calico Life Sciences LLC, South San Francisco, California; and also attended the Walter and Eliza Hall Institute of Medical Research and the University of Melbourne, both in Australia.

The research was supported by the Texas Cancer Research Research Institute (RR160057) and the National Institutes of Health (T32CA080621; F32GM117793; K25CA204526, R33CA235254, and R35GM133522). Fiolka has filed a patent for the scanning unit and its applications in microscopy. There are additional disclosures in the newspaper. The researchers made the data used in the study and the software code available online. Access details are in the studio.

Fiolka is a member of the Harold C. Simmons Comprehensive Cancer Center. Visit the Fiolka lab here.

About UT Southwestern Medical Center

UT Southwestern, one of the most important academic medical centers in the country, integrates pioneering biomedical research with exceptional clinical care and education. The institution’s faculty has received six Nobel Prizes and includes 24 members from the National Academy of Sciences, 16 members from the National Academy of Medicine and 13 researchers from the Howard Hughes Medical Institute. The workforce of more than 2,800 full-time professors is responsible for innovative medical advances and is committed to quickly translating science-based research into new clinical treatments. UT Southwestern physicians provide care in nearly 80 specialties to more than 117,000 inpatients, more than 360,000 emergencies, and monitor nearly 3 million outpatient visits a year.

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