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Taking a Closer Look at Trends in Microscopy

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Imaging has revealed many secrets that are hidden to the naked eye, giving enlightenment to scientists, and helping to solve biological conundrums. Without microscopy capabilities, the causes and solutions to many disease and conditions that are now treatable would remain a mystery. With advances in cellular and tissue-based imaging, researchers are able to peel back the layers on new challenges and test hypotheses to solve more problems. However, for them to be able to do this, they require the tools with which to do it. It is therefore vital that the technology behind microscopic imaging systems continues to advance at the same pace.

We spoke to Dr Christine Munz, Vice President of Life Science at Leica, about progress being made in the microscopy field, and how advances in confocal microscopy are changing the imaging landscape for scientists.

Karen Steward (KS): What do you see as the biggest driving forces behind advancements and R&D within the field of microscopy?

Christine Munz (CM): The driving force behind advancements in the field of microscopy is the need to continuously support our customers, scientists. They are constantly looking for ways to move beyond current limitations concerning observation and detection. Why? Because they want to take the next step and be able to achieve a better understanding of the true nature of the biological specimens or processes they explore. The key to the growth of modern microscopy is the evolving ability to clearly and accurately visualize specimens in three dimensions. Extending the usable spectral range, like we have done with the STELLARIS confocal platform, allows more fluorescent labels to be observed simultaneously without making the experimental setup more complex. Keep in mind that every increase in detection efficiency pushes back the detection limits that much further. In a similar vein, instrument manufacturers are developing solutions for extracting as much information as possible from specimens. By taking advantage of fluorescence lifetime information, it is even possible to add a new “dimension of information”. As it is fast and fully integrated into the detection system, fluorescence lifetime provides immediate access to functional information that is simply not available in conventional confocal systems based on intensity detection only. This scientific need has driven the innovations now implemented in the STELLARIS platform. The performance of individual components, such as detectors and lasers, were taken to the next level and, with lifetime-based imaging, new dimensions of information are made accessible to every STELLARIS user. With the STELLARIS platform, scientists can see more and discover more or, in other words, get closer to the truth.

KS: Why is it important to be able to capture 3D images of living cells and tissues? What important insights can be obtained that 2D imaging lacks?

Information in 2D can be extremely valuable, depending on the scientific question that is being asked. However, cells and multi-cellular structures are 3D objects. The focus of today’s science shifts towards understanding the functional processes in the cellular context, typically from highly valuable specimens. Therefore, it is important to enable not just the exploration of the three-dimensional space, but also the maximizing of information that can be extracted, e.g. through extended multiplexing.

KS: What are some of the challenges faced in improving confocal microscopy?

A central challenge is to get by with as little light as possible to protect sensitive cells and specimens while, at the same time, obtaining a maximum signal to produce the most meaningful images possible. In order to make as many structures visible at the same time as possible with different dyes, microscope systems must cover a wider range of visible light than ever before. This need requires a complete revision of the entire beam path and detectors. In addition, researchers are increasingly expected to correlate their fluorescence images with other data. This goal requires that information beyond purely light intensity must be readily accessible. Making the so-called fluorescence lifetime available for this purpose required a fundamentally new concept of data acquisition in Leica confocal laser scanning microscopy (CLSM).

KS: Many analytical areas are moving towards equipment that can be operated by non-expert users. Is the field of confocal microscopy following the same trend?

We see many scientists utilizing a multitude of technologies to answer complex biological questions. Therefore, on the one hand we need to further simplify the way microscopes are operated, but at the same time we need to expand the insights they deliver. Therefore, we have designed the STELLARIS platform towards simplicity, even for complex experiments, and to help users identify relevant details instantly, while also delivering the power to see more and the potential to discover more.

KS: Can you tell us about any specific cases where 3D confocal imaging has been a real game changer in research?

There is a recent publication related to host-pathogen interaction in a newly discovered immune disease. The researchers imaged cells with and without a mutation for an important zinc channel that is involved in a new immunodeficiency. The LAS X Navigator (a software module for multiparametric acquisition and data review of large data sets) with the FALCON system (FAst Lifetime CONtrast) allowed us to see that, with the mutation, there is no longer a zinc gradient in the cells and this causes the diminished immunity in the patients.

Dr Christine Munz was speaking to Dr Karen Steward, Science Writer for Technology Networks.