A new method of microscopy provides a look into the future of cell biology – ScienceDaily

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?

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, could open new avenues in advanced microscopy, the researchers say.

“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 holding the biological specimen with your hand, turning it over and inspecting it, which is an incredibly intuitive way to interact with a sample. By 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 customized 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, ignoring 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 may represent a new paradigm for acquiring information in 3D using a fluorescence microscope.”

It also promises incredibly fast images. While an entire 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 moment! We realized that this could be bigger than a simple optical deflection method; that the system could also work for other types of microscopes,” 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.

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