In the world of super-resolution fluorescence microscopy, MINFLUX is providing a new level of detail to molecular biology pursuits. <\/p>\n\n\n\n
It can resolve biomolecules only nanometres apart. It can even capture rapid biological movements at this very discrete level, such as molecules moving within a living E. coli<\/em> bacterium. But something this powerful microscope can not<\/em> do is train scientists to harness its incredible capability. <\/p>\n\n\n\n
That\u2019s where EMBL and the EMBL Imaging Centre step in.<\/p>\n\n\n\n
Home to some of Europe\u2019s foremost cutting-edge tools in molecular biology, EMBL has long shared its expertise and access to these tools through an extensive repertoire of courses, conferences, seminars, and other training. And now included in this mix is a job shadowing programme at the EMBL Imaging Centre (EMBL IC) aimed at \u2018training the trainers\u2019.<\/p>\n\n\n\n
While academic papers and conferences can offer a window into the latest scientific technologies, the EMBL IC job shadowing programme provides up to three months\u2019 stay at EMBL, with a mix of observation, consultation, and hands-on experience.<\/p>\n\n\n\n
\u201cRight now, there are only a handful of facility MINFLUX systems in the world,\u201d said Virginia Pierini, EMBL IC service manager, noting how other instrumentation and expertise also attract scientists to this individualised training approach. \u201cWe are in a unique position to share a variety of expertise on some of the very best microscopes, but also techniques that have been honed here at EMBL and workflows that might improve services elsewhere.\u201d<\/p>\n\n\n\n
After the EMBL IC opened in 2022, the staff provided this kind of advanced training to four visitors who came from Sweden, Switzerland, the United States, and Singapore. The programme continues on in 2023, overseen by Pierini, and with Zhengyi Yang, an EMBL cryo-electron tomography specialist, serving as guide, trainer, and liaison for visitors to meet other EMBL experts. <\/p>\n\n\n\n
\u201cJob shadowing and this intensive training of staff who come from other international imaging facilities and large labs are part of a programme that aims to accelerate the use of new service workflows and new imaging technologies,\u201d Pierini said. \u201cThe training we offer is unique because it focuses on correlative technologies, meaning we use several different methods of light or electron microscopy to analyse the same cell or tissue and combine these different perspectives.\u201d <\/p>\n\n\n\n
Maxing the MINFLUX knowledge<\/strong><\/h2>\n\n\n\n
For Hans Blom, Head Staff Scientist at the Advanced Light Microscopy facility at Sweden\u2019s National Life Sciences Laboratory (SciLifeLab), a visit to EMBL in 2022 meant he could get hands-on experience with MINFLUX, which his facility then purchased this year. <\/p>\n\n\n\n
\u201cIt was only two days in November when I pursued my MINFLUX visit, but having that one-on-one training by an expert (and very long days) \u2013 it was effectively like a normal weeklong course,\u201d Blom recalled.<\/p>\n\n\n\n
Part of Blom\u2019s team specialises in super-resolution fluorescence microscopy support for national and international user projects. SciLifeLab \u2013 in conjunction with his facility\u2019s academic host, the Royal Institute of Technology (KTH) \u2013 had provided funding beforehand to buy a MINFLUX platform to complement and extend its other super-resolution microscopy systems. This is why Blom wanted to learn more specifically about the MINFLUX technique and get preliminary training on the system while receiving tips for conditions to help it work optimally when it arrived at his own facility. <\/p>\n\n\n\nAs head of the Advanced Light Microscopy facility at Sweden\u2019s National Life Sciences Laboratory (SciLifeLab), Hans Blom spent just two days at EMBL Imaging Centre learning about MINFLUX, but described the experience as necessary to get the augmented know-how and skills to be prepared to do a good service job. Credit: \u200b\u200bHjalmar Brismar SciLifeLab\/KTH<\/figcaption><\/figure>\n\n\n\n
Blom had heard about the training opportunity through Timo Zimmermann, who heads up the light microscopy facility within the EMBL IC. <\/p>\n\n\n\n
\u201cIn the last few years, I have followed the development of MINFLUX. I have been interacting with research colleagues in both Sweden (KTH) and Germany (University of Jena and University of G\u00f6ttingen), who are applying and developing the MINFLUX technique further.\u201d Blom said. \u201cThis research interaction is great, but to do the harder task of providing external support with advanced techniques, train-the-trainer education is crucial. Facility support with any advanced technique demands consistency of support on all levels to many non-experienced users. Only by learning from experienced facility staff experts does one get the necessary augmented know-how and skills to be prepared to do a good service job.\u201d<\/p>\n\n\n\n
As a facility head, Blom follows most international imaging facilities and networks to interact as much as possible with fellow staff scientists, noting that this kind of engagement is crucial to continuously advance support for external users. <\/p>\n\n\n\n
\u201cAny job shadowing or train-the-trainer initiative is extremely helpful,\u201d he said. \u201cIt both raises the quality of support as well as democratises its use.\u201d <\/p>\n\n\n\n
Expanding the range of methodologies<\/strong><\/h2>\n\n\n\n
While MINFLUX is definitely one of the newer imaging technologies, other scientists like Thorsten Blum have also come to EMBL, focusing on a broad range of cryo-electron microscopy (cryo-EM) methods.<\/p>\n\n\n\n
A cryo-electron microscopist in the Volodymyr Korkhov group at the Paul Scherrer Institute (PSI) in Switzerland, Blum\u2019s EMBL experiences of learning and experimenting with state-of-the-art techniques will also benefit other scientists at PSI. He has been at EMBL for the past three months.<\/p>\n\n\n\n
\u201cTo be able to learn about the various methods by literally looking over the shoulders of experts and even being able to do my own processing with the help of experienced colleagues, sounded very appealing to me,\u201d said Blum. <\/p>\n\n\n\n
He has been deepening his knowledge in sample preparation in two different ways: writing patterns for cell arrangement on electron microscopy grids and focused ion beam milling. Ion beam milling produces thin slices of cells that are hundreds of nanometres in thickness. It allows researchers to achieve sub-nanometre resolution in their imaging and to better observe cells in their native states when using transmission electron microscopy. <\/p>\n\n\n\n
Blum\u2019s experiences at EMBL also have focused a lot on data acquisition workflows and different ways to make sure the data obtained are high quality as well as high throughput. This was supplemented by seeing data processing strategy options employed in different research groups, particularly those of Simone Mattei and Julia Mahamid, EMBL Team Leader and Group Leader, respectively. He has also been able to try out some of the newest data processing software at EMBL. <\/p>\n\n\n\n
\u201cEMBL Imaging Centre, other facilities, and labs are impressive on their own, having some of the latest technologies,\u201d Blum said. \u201cAdditionally, there have been demos and visits from engineers from the centre\u2019s industry partners [Leica Microsystems, ThermoFisher Scientific, Carl Zeiss Microscopy, Abberior Instruments, CryoSol-World]<\/em> who provided useful discussions. So, the knowledge transfer is impressive on both sides.\u201d <\/p>\n\n\n\n
Blum looks forward to applying new techniques to his work at PSI, such as the high-pressure freezing protocol for sample preparation. <\/p>\n\n\n\n
\u201cI want to test a new protocol for data collection on our microscopes, and after my stay, I will have a pipeline for subtomogram processing. I am now quite eager to install on our cluster the software I\u2019ve used here,\u201d he said. \u201cBefore coming, I had imagined EMBL to be at a very high standard. In reality, it turned out to be even more impressive, providing a way to return home with ways to implement a framework for projects that can be adapted for specific needs.\u201d<\/p>\n\n\n\n
Ann Tran Bich Ngoc was EMBL\u2019s first participant in the job-shadowing programme \u2013 a testament to the EMBL Imaging Centre\u2019s worldwide reputation as the place to learn about cutting-edge imaging. Coming from Nanyang Technological University, Singapore, she is involved in managing the cryo-EM facility there and provides training in techniques related to sample preparation and imaging. <\/p>\n\n\n\n
\u201cEMBL Imaging Centre houses a full range of state-of-the-art cryo-EM equipment and highly capable scientific staff \u2013 a place where I could accelerate my learning in both instrument operation and setup\/maintenance, which certainly benefits our users,\u201d she said.<\/p>\n\n\n\nAnn Tran Bich Ngoc, from Nanyang Technological University, Singapore,\nwas EMBL\u2019s first participant in the EMBL Imaging Centre\u2019s job-shadowing training. Credit: Kinga Lubowiecka\/EMBL<\/figcaption><\/figure>\n\n\n\n
A key training goal was to explore alternative workflows that might help Ngoc manage her facility more efficiently.\u00a0 \u201cWe’ve now applied some technical advice I got at EMBL into our experiments,\u201d she said. \u201cLikewise, some workflow suggestions have also been helpful, especially with respect to sample preparation, data acquisition, and data processing for single particles and tomography.\u201d<\/p>\n\n\n\n
Ngoc added, \u201cby following Anna Steyer (cryo-tomography specialist) and Zhengyi Yang as they did their experiments, and engaging in them, I really could learn more about how to handle different types of samples, optimise sample preparations, acquire good data, and engage in troubleshooting.\u201d<\/p>\n\n\n\n
Yang, who has guided two of these visiting scientists\u2019 experiences, sees the programme as being useful to EMBL in many ways. <\/p>\n\n\n\n
\u201cThe experience provides for a fruitful information exchange. I really enjoy being involved in it,\u201d Yang said. \u201cThe knowledge exchange benefits both sides. We also get to learn how the workflow is implemented in other institutes. All in all, it\u2019s the kind of exchange that just makes the scientific process better.\u201d<\/p>\n\n\n\n
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