Taking the Potential of Confocal Microscopy across the Diffraction Barrier
Confocal microscopy, which enables scientists to construct 3D representations of objects in the range of hundreds of nanometer, has been the workhorse and method of choice for imaging for decades now and has revolutionized our view of biology and life sciences. Scientists at VIB have taken the potential of confocal microscopy across the diffraction barrier – using innovative software-based image analysis tools. In the past, it was believed that the laws of physics make it impossible for classical confocal microscopy to resolve anything smaller than 200 nanometers. This new, patented methodology now makes this possible and will be further developed through a collaboration between VIB and German microscopy company Zeiss.
The 200-nanometer diffraction barrier has been considered a physical limit that traditional microscopy could not be expected to cross. Below this barrier, it is impossible to determine the difference between two points using the classical microscopy technology available in many labs. Although in the recent past super-resolution techniques have been developed, they have been limited, as samples were tedious to prepare and/or needed new expensive hardware. With the new development, prof. Sebastian Munck and his VIB BioImaging Core are taking confocal microscopy to the next level, as the standard off-the-shelf devices can be used with no need for additional hardware or complicated sample preparation.
Spurred by the never-ending need for more data to validate scientific research, VIB researchers have developed a completely new and innovative technique that enables super-resolution with confocal microscopes.
The best of two worlds: super-resolution confocal microscopy without the hassle
Scientists at VIB are well-positioned to be familiar with demands and pain points in microscopy. Some of the hassle involved with other super-resolution techniques include complex sample preparation, optimizing protein labeling, and a limited number of colors channels. Originally titled Point Detection Imaging Microscopy through Photobleaching, VIB’s new technique increases resolution in biological samples beyond the diffraction limit and makes 3D imaging possible.
One of the great advantages is the versatility of the approach, by making it possible to apply super-resolution on off-the-shelf microscopes. This means that all the benefits and decades of optimization like standard sample preparation and customer oriented developments like multicolor imaging are ready to be used.
Prof. Sebastian Munck (VIB BioImaging Core Leuven): “We wanted to make super-resolution super easy to be able to focus on our research questions instead of focussing on the technology.”
Making this technology available for the entire research community
After inventing the technology, the next step was to engage with industry. Germany base microscopy company Zeiss is charging their confocals with super-resolution powers based on VIB technology.
Dr. Ralf Engelmann (Product Manager 3D Microscopy, Zeiss): “Superresolution is still one of the main trends in microscopy and a strong driver for new discoveries. SR-DIP, as we will call the new module for our ZEN blue software platform, makes superresolution available for researchers who don’t want to do big hardware investments, but still want to do cutting edge research.”
Dr. Jérôme Van Biervliet (Senior Business Development Manager, VIB): “We are excited that this technology will be available for research via the commitment of Zeiss. This is a tool for even better research, enabling even more thorough molecular understanding of how molecular players in disease functionally interact at disease onset and progression – with the ultimate end goal of developing even more effective interventions.”