New imaging systems help scientists look deeper into cells – and materials, too
Applications of machine vision extend beyond industry: Some exciting uses are found in scientific research, from the life sciences and even medicine to materials science.
BioPhotonics consulted some experts in the field to find out the current trends in vision for scientific research – and what the future holds. They included Max Larin, CEO of Ximea in Münster; Dr. Tanjef Szellas, product management leader at Leica Microsystems in Mannheim; and Markus Wiederspahn, senior manager of communications at Carl Zeiss Microscopy in Oberkochen, all in Germany. The three companies offer imaging equipment for scientific applications.
Cameras for scientific research allow samples to be viewed with ever-improving resolution. Here, a volume rendering from the results of reconstruction of a wood sample scanned at 500-nm spatial resolution. Courtesy of Ximea GmbH.
Q. How would you say the market has been in the past few years for scientific vision systems?
Szellas: There was a clear growth in confocal microscopy, especially for advanced systems. People now want to purchase systems that can do a wide range of applications – this is part of what we have been seeing in the past 10 years toward consolidation of equipment between research groups, or the setting up of centralized equipment parks (core facilities).
Wiederspahn: The market has seen some growth in the past years, but it has now become flat due to limits in public budgets. Industry research is only partially covering this shortfall.
Larin: Technological changes are coming faster and with less predictability. This can be a challenge for system developers but is extremely exciting for end users – namely scientists, engineers and technicians – because of the variety and power of new imaging tools and what they mean for cutting-edge scientific research and medical care/diagnostics.
Q. Where do you think the market is going?
Larin: While products may be coming to market faster than ever before, funding for research and hardware means manufacturers need to get used to shrinking margins and increasing competition.
Szellas: The trend from purely biologically oriented research use toward medically relevant research (diagnostics) is likely to continue. Basic research into lifestyle-related diseases is going to be even more prominent than before – and microscopy is at the heart of most of that basic research.
This screen shot of three orthogonal virtual slices show individual cells inside a wood sample nondestructively visualized by a micro-CT system with 500-nm isotropic resolution. Courtesy of Ximea GmbH.
Wiederspahn: There are huge challenges in life sciences as well as in materials sciences. Intriguing tasks like brain research/connectomics, cancer root cause analysis, embryology or graphene research, nanofabrication – to mention but a few – require by far more instrumentation than is existing nowadays.
Q. Which application areas are thriving, and why? Which is a “bigger” area, life sciences applications or materials science applications?
Szellas: For Leica, life sciences research is definitely the currently more relevant field. Materials science is more affected by economic factors so can rise and fall quite quickly.
Larin: Ximea saw more activity in materials science during the past year, but we wouldn’t say that is a global trend based solely on our observations. Materials science, outside of large industries such as semiconductors that use very specific tool sets, is still a relatively small area of development compared to life sciences and medical care.
Wiederspahn: Live-cell imaging, e.g., observation of cells in a physiological environment ... will become more and more mainstream and is, therefore, thriving.
A confocal image of a mouse retina, reproduced with permission from Dr. Frank Müller of the Institute of Complex Systems-Cellular Biophysics at Research Center Jülich GmbH in Jülich, Germany. Courtesy of Leica Microsystems.
However, also many materials science applications – e.g., graphene research or nanofabrication/nanomodification – offer lots of opportunities to us.
One major trend in life sciences is derived from the capability to get high-resolution images using field-emission scanning electron microscopes which hitherto were only to be obtained by transmission electron microscopes. This, together with correlative light and electron microscopy and serial block-face imaging, are upcoming trends to be found.
Q. What do you see as the “next big thing” in scientific imaging in general?
Larin: Back-side illuminated sensors, sensors that improve quantum efficiency for low-noise, low-light and ultrafast imaging applications, coupled with fast multigigabit back-ends for highest possible throughput for scientific imaging.
Szellas: There is a clear trend from purely descriptive techniques toward quantitative imaging. This is combined with all technologies used in confocal microscopy: single-molecule detection, live-cell imaging, superresolution, etc.
Wiederspahn: Selective plain illumination microscopy (SPIM) or light-sheet fluorescence microscopy will become a major technology in scientific imaging of living organisms and plants, enabling observation of samples over a period of several days in physiological conditions and ... videos of developments of samples opens up completely new opportunities to researchers. Carl Zeiss holds several patents on this technology and is developing a product.
Q. Are you seeing any new and exciting scientific imaging advances coming out of R&D and/or university labs?
Szellas: Apart from forward steps in technology, superresolution is definitely the field with the most obvious imaging advancement in microscopy within the past five to 10 years – and it’s still progressing, fast! There is exciting research going on with respect to dye development that synergizes with advanced imaging techniques.
Neurobiologists now can capture wide-field images of synapses at the neuromuscular junction of drosophila, as shown here with triple staining for the presynaptic active zone marker Brp (green), postsynaptic glutamate receptors (red) and the presynaptic membrane (blue). Image by Jan Pielage of Friedrich Miescher Institute in Basel, Switzerland; Provided by Carl Zeiss Microscopy.
Other areas that are rapidly advancing are: screening (both high-content and high-throughput), and label-free imaging (CARS), shifting from pure physics toward biomedical research.
Larin: Sony stacked CMOS sensors.
Wiederspahn: 3-D imaging of (especially) biological samples with voxel sizes in the nanometer range becomes possible by methods such as 3-D block-face imaging or the combination of milling and imaging using combined focused ion beam/scanning electron microscopes such as the AURIGA CrossBeam workstation. A major application of these methods is in brain research, namely connectomics. For the first time researchers using brain mapping are getting closer to understanding the neuronal network in the brain. But also, other organs can be examined using these methods, again opening up new insights in structure and functionality.
Q. What are the biggest challenges to new advances in scientific imaging?
Szellas: Data management: Huge amounts of data need to be transferred from microscope to server to analysis to automated interpretation. Limitations by dyes (e.g., size of fluorophores in superresolution), and their suitability for live-cell imaging.
Wiederspahn: There are at least three main challenges to be mentioned.
All new imaging technologies deliver enormous amounts of digital data. Biologists or materials scientists are faced with the challenge to store these data and to do image processing as well as identify specific areas. New approaches like gamification help to leverage the crowd as a source of knowledge.
Automation: Applications like connectomics, research using correlative microscopy, serial block-face imaging will require a high degree of automation in order to save valuable time.
Sample preparation work flows: With all the new methods evolving, improved and automated sample preparation work flows need to be available.
Larin: For camera makers, it is to keep control over development time and production cost to capitalize first mover advantage of important camera/system improvements.
For scientists, doctors and technicians, it is how to acquire funding for the latest tools to take your research or patient care to the next level.
Scientific imaging equipment
Ximea, Leica and Carl Zeiss all offer vision tools for scientific research, designed for both the life sciences and materials science, and their offerings clearly illuminate the trends in these fields.
The xiRAY11 from Ximea GmbH.
Ximea’s xiRAY11 is an 11-megapixel, fiber-coupled and cooled x-ray camera based on Truesense’s KAI-11002 sensor. The camera features Ximea’s sensor driving technology, Cleanpath, which enables it to deliver crystal-clear 14-bit images with a 36 x 24-mm field of view. It has a 4-fps refresh rate in full resolution mode (12 fps with 4 x 4 binning), as well as programmable exposure times from 12 µs to 500 s. “All these features come in a camera module measuring only 63 x 63 x 40 mm,” Larin noted.
These properties provide unprecedented resolution, speed and performance to Bruker microCT’s new micro-CT scanners, compared with other x-ray camera approaches to intermediate-resolution scans, Larin said.
The TCS SP8 from Leica Microsystems.
The Leica TCS SP8 has a 12-kHz scanner for scanning up to 430 fps as well as GalvoFlow for fast 3-D recordings, Szellas said. Its HyD detectors (descanned and nondescanned) enable live-cell imaging with minimal photodosage and high contrast.
The camera’s specially designed scan optics benefit science applications, he said, as does the all-new software interface with application wizards and advanced 3-D visualization.
The TCS SP8 offers superresolution through gated stimulated emission depletion technology for sub-50-nm lateral resolution in combination with a white-light laser. It also can be used for label-free imaging based on vibrational contrast as in coherent anti-Stokes Raman spectroscopy as well as high-content screening.
The Elyra from Carl Zeiss Microscopy.
Addressing the trend toward higher-resolution imaging and superresolution microscopy, Carl Zeiss has combined two superresolution methods into a turnkey platform called Elyra. The system can be configured for superresolution structured illumination, photoactivated localization microscopy, or both at once. It also can be combined with a laser scanning microscope, Wiederspahn said.