74 An international fi rst
Beyond the microscopy and scientifi c software experts however, it is primarily the researchers themselves that drive and guide developments in imaging technology. Beamline B24 has been privileged to work with enthusiastic and committed research scientists that engaged with the methods, fed back to the design process and drove automation development. As a result, the correlative imaging platform at B24 can now deliver an international fi rst in user-friendly, high-resolution, 3D correlative cryo-imaging. As Dr Stephen Graham, a Sir Henry Dale Fellow and University Lecturer at the Division of Virology of the University of Cambridge commentedz, “The facilities at B24 give us unique tools to probe virus infection in cells. By correlating super-res light cryo-microscopy with soft X-ray cryo-tomography we can understand exactly what stage of infection each cell is at and can probe the structural rearrangements that viruses induce in the host cells that they exploit. The fi xation-free and stain-free workfl ow plus the huge depth of fi eld available using cryoSXT is especially advantageous for us as it allows us to capture rare events within the cell, like catching viruses in the act as they cross the nuclear membrane.”
Dr G Tozzi’s team collecting data at B24 on the morphology of differentiated stem cells from sheep.
Commercial interest in correlative microscopy at B24 has been growing fast and recent client projects have focused on key areas for the pharmaceutical industry such as novel therapeutics and vaccine development. Of particular value to industrial clients is the beamline’s full service approach, where no prior knowledge is required. The expert team at B24 can support all stages of the project, from experimental design and data collection through to analysis and reporting. Our clients greatly value the scientifi c insight, gained in a timely manner, which feeds into their own research and development pipelines.“
Getting the message across Dr. S Graham and his collaborator Dr C Crump collecting data on HSV1 infected cells at beamline B24
Dr Gianlucca Tozzi, a reader in Bioengineering and Director of the Zeiss Global Centre at the University of Portsmouth in the UK has used the B24 imaging platform in his studies on biological tissues and biomaterials for tissue engineering. He said: “Working at B24 has been a great experience. Support in all technical aspects has been brilliant and there is a real will to push further development of the technology in addressing complex imaging modalities, which are increasingly needed to unravel biological mechanisms. I am looking forward to continuing this excellent collaboration”
This novel imaging approach is particularly relevant to the fi eld of cancer biology. As Professor Peter Saddler from the Chemistry Department at Warwick University in the UK highlighted: “B24 has added new dimensions to our work on elucidating the mechanisms of action of novel anticancer drugs which can combat resistance to treatment, in particular prodrugs which are activated specifi cally in cancer cells by visible light. Correlative super- resolution fl uorescence microscopy and 3D soft X-ray tomography allows us to monitor both the localisation of fl uorescent drugs and drug-induced subcellular changes to organelles, such as mitochondria and endosomes, in cryopreserved cancer cells close to their native state.”
Signifi cantly, industry also acts as a driving force for development at the beamline. As Dr Claire Pizzey, deputy head of Industrial Liaison at Diamond put it: “Industrial engagement is an important aspect of the work of any beamline at Diamond given that up to 10% of operational time can be dedicated to proprietary use for confi dential research and development.
Often and unexpectedly the major hurdle once a microscopy setup is operational is to let the people that will most benefi t from it know that it is now available and how they can take advantage of it in the most time-effi cient and productive way. At B24 processes are fully documented on our websites (
www.diamond.ac.uk/Users.html and https://www.diamond.
ac.uk/Instruments/Biological-Cryo-Imaging/B24.html) and accompanied with protocols and expertise that allow the standardisation of the steps required and remove the element of guesswork from sample preparation and processing as well as data collection and analyses.
At B24, the development of a new imaging platform has fi lled previously unmet needs of biological imaging by combining the power of cryo-SIM and cryo-SXT microscopes to provide valuable insights into the inner workings of biological material. One method allows us to map the location of chemical signatures and the other gives us ultrastructural information and cellular content, literally a CT scan of a cell with highlighted areas of interest. Once together the researcher has both a 3D map of a cell, and the specifi c location within it of molecules of interest. This innovative and largely user-friendly microscopy setup at beamline B24 perfectly demonstrates what can be achieved by multidisciplinary collaboration and is now available to the wider research community.
Further information from
maria.harkiolaki@diamond.ac.uk References
1. Kounatidis, I. et al. 3D Correlative Cryo-Structured Illumination Fluorescence and Soft X-ray Microscopy Elucidates Reovirus Intracellular Release Pathway. Cell 182, 1–16 (2020).
Read, Share and Comment on this Article, visit:
www.labmate-online.com/article New Scanning Electron Microscope with Automated Analytical Intelligence
Jeol introduces a new Field Emission Scanning Electron Microscope with several features unique to the company’s FE SEM product line: NeoEngine, employing analytical intelligence for optimising electron beam setup and tuning; embedded EDS with Live Analysis for real time imaging and elemental analysis; and Zeromag navigation function, seamlessly transitioning between optical imaging to nanoscale investigation with the high-powered optics of the SEM.
“Jeol’s F100 FE-SEM offers a truly revolutionary approach to address users’ high-resolution microscopy and microanalysis needs. In a single platform Jeol combined the best electron optics with fully embedded EDS microanalysis and the powerful AI algorithms of NeoEngine to achieve the ultimate ease of use and streamlined workfl ow,” said Dr Natasha Erdman, FESEM Product Manager.
NeoEngine corrects electron trajectories and automatically aligns the beam in real time, plus automatically corrects focus, brightness/contrast, and astigmatism. The SEM achieves single-digit Angstrom resolution at low kV, ideal for ultrahigh high-resolution imaging of nanostructures, specimen surface details, biological specimens, and magnetic samples as well as elemental analysis of non-conductive and beam sensitive samples. Fully embedded EDS with Live Analysis allows the user to simply select the area, mapping, line, or another type of analysis directly on the observation screen to begin automatic live display of the elements in the specifi ed location.
The large specimen chamber of the JSM-F100 features multiple ports for analytical applications: EDS, WDS, EBSD, STEM, BSE, and CL. An optional Soft X-ray Emission Spectrometer provides effi cient and parallel collection of very low-energy X-rays while providing unprecedented chemical state analysis. The Jeol JSM-F100 features an in-lens detector and energy fi lter, an Aperture Angle Control Lens (ACL) for superb resolution at any kV or probe current, Beam Deceleration (BD) mode for charge reduction and enhanced surface detail, and a variable pressure option for non-conductive specimens. Never before has the power of Field Emission SEM been so easy to use. With the introduction of Jeol’s next-generation streamlined FE SEM product line, operator controls are simplifi ed and the functions of the SEM fully automated for optimum performance. Imaging and analysis data are quickly obtained and reported through the effortless data management system.
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INTERNATIONAL LABMATE - SEPTEMBER 2020
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