Instrumentation and Technology updates from Instruct Centre CZ


CIISB, an Instruct-CZ Centre,offers priority access to groups that need to use CIISB structural biology services for projects directly related to studies of the SARS-CoV-2 virus and projects aiming to develop an effective vaccine or treatment. Successfully accepted proposals are free of charge, and no financial contribution is requested for the measurement/service. To request priority access, a research proposal with „COVID-19“in the title of the proposal should be submitted through the online application system. In the past, CIISB services operated mostly with users attending at or running their experiments. However, under this year’s pandemic measures the remote operation/data collection has been rather popular, reaching on average about 40-50% of all the provided experimental time. For the individual core facilities, the amount of remote access varies, ranging between 15% (Diffraction Techniques) and 100% (Proteomics).


Instrumentation and Technology updates


The Biomolecular Interactions and Crystallization core facility installed a new Bio-layer interferometer (BLI) Octet RED96e for analysis of macromolecular interactions. Automated differential scanning calorimeter PEAQ-DSC was acquired foranalysis of protein structural changes upon heating. The addition of Dynamic light scattering (DLS) plate reader Spectrolight600 increased capacity forhigh-throughput detection of aggregation and sample stability. Acquisition of multidetector-coupled size-exclusion chromatograph (SEC-MALS) OmniSEC serves for detailed analysis of sample homogeneity and size of molecules and their complexes. Till the end of 2020,full operationof new crystallization storages shall be finished.

Cryo-electron Microscopy and Tomography core facility acquired the high-pressure freezer Leica EM ICE for vitrification of bulky biological specimen (up to 200um thickness). In addition, the freeze-substitution unit Leica EM AFS2 for resin embedding of the high-pressure frozen samples and the ultramicrotom Leica EM UC7 with the adapter for cryo-ultramicrotomy were purchased to provide the facility users with the complete workflow for preparation of thin section samples for both room-temperature electron microscopy and cryo-electron microscopy. In addition, electron diffraction tomography (micro-ED) was implemented as a standard service provided by the facility and a new detector Dectris Quadro was installed to increase quality and throughput of micro-ED data.

The Crystallization of Proteins and Nucleic Acids core facility extended automated screening to low temperatures with a new crystallization hotel RI182 (Formulatrix) to broaden the already available options of dedicated spaces for crystallization at three different temperatures. After a survey to map the demand for a new facility focused on protein production services, a small protein production unit was formed, currently in pilot operation, to evaluate its feasibility for the long run. In the Josef Dadok National NMR Centre,the exchange of electronics of all high-field spectrometers(600, 700, 700, 850, and 950 MHz)to Avance Neo (Bruker) was completed.The four systems with cryoprobes were equipped with nitrogen liquefiers that extend the service interval from 2-3 weeks to more than 6 months.

The Nanobiotechnology core facility equipment was extended with a combined system of Renishaw InVia Raman microscope and AFM microscope Bruker Icon to combine high resolution imaging with ability of chemical mapping and imaging. The JPK NanoWizard3 AFM was upgraded with high speed and sensitives CMOS camera And or Zyla 5.5 integrated fully to the JPK software, thus allowing combined online fluorescence and AFM imaging.

In the Proteomics core facility,the first hybrid mass spectrometer with ion mobility module timsTOF Pro (Bruker) was installed. Additional separation dimension coupled with advanced acquisition modes increase our capabilities for deeper qualitative and quantitative characterization of complex protein samples.

The core facility Structural mass spectrometry introduced a new mass spectrometer TimsTOF (Bruker Daltonics). The mass spectrometer will be used for high-throughput proteomics approaches, ion mobility separation of molecules and other structural analysis of proteins including hydrogen-deuterium exchange, chemical cross-linking and covalent labelling.

The X-ray Diffraction and Bio-SAXS core facility upgraded the goniometer (from geometry partial chi to kappa-geometry) and the X-ray detector(from CCD to HPC, Hybrid Photon Counting detector) at the dual wavelength Rigaku single crystal diffractometer.

New mass spectrometer timsTOF Pro was installed in November 2020 in the Structural mass spectrometry core facility in BIOCEV and is now fully in operation. The mass spectrometer will be used mostly for shotgun proteomics, hydrogen-deuterium exchange and covalent labelling experiments, and native mass spectrometry with ion mobility separation. The same system was installed also in the Proteomics core facility at CEITEC.