CeNT
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Cryomicroscopy and Electron Diffraction Core Facility

Tomasz Góral, PhD - experiment design consultant, microscope operator and data analyst

Research qualifications and technical skills:

  • Expertise in sample preparation for cryo-EM including glow discharging, plunge freezing and autogrid clipping

  • Fluency in operation of the 200kV cryo-TEM Glacios including EPU and TOMO software packages

  • Fluency in operation and calibration of a range of scanning electron microscopes: Zeiss Ultra Plus, FEI QUANTA 650 (ThermoFisher brand), LEO 1455VP, Phillips XL30, Zeiss EVO 15LS and a transmission electron microscope Hitachi 7100

  • Fluency in EDS analysis of a range of different samples both biological and mineralogical

  • Fluency in sample preparation techniques for SEM including dehydration and critical point drying. Experience in using high resolution sputter coaters

  • Confocal microscopy with expertise in fluorescence recovery after photobleaching (FRAP) technique, working knowledge of fluorescence lifetime imaging (FLIM) technique and total internal reflection (TIRF) microscopy. Fluency in operation of the Nikon A1- Si and Zeiss LSM700 confocal microscopes

  • Confocal image processing including signal deconvolution

  • Working knowledge of X-ray nano-computed tomography (XuM) and micro-CT imaging

  • Working knowledge of the Alicona Infinite Focus microscope

  • Operational knowledge of several imaging softwares: NIS-Elements (Nikon), Zen (Zeiss), LAS (Leica), Volocity 3D Image Analysis (Perkin Elmer), Imaris (Bitplane), Image Pro (Media Cybernetics), Volume exploration and presentation tool – Drishti, MeX (Alicona), ImageJ
Kunal Kumar Jha, PhD – experimental design consultant, microscope operator and data analyst

Research qualifications and technical skills:

  • Expertise in sample preparation for 3DED measurement

  • Expertise in data collection strategy and structure solution of micro-nano crystals of small molecules from X-ray and electron diffraction measurement

  • Operational knowledge of the 200KV Cryo-TEM Glacious with EPU-D software package

  • Expertise in operation of Single crystal X-ray diffractometer BRUKER D8 Venture with APEX II,III,IV software package

  • Expertise in crystal growth using various methods for generation of polymorphs

  • Expertise in processing and structure solution of complex single-crystal X-ray and electron diffraction data having different types of disorder in organic small molecule, organometallic compounds etc.

  • Expertise in High-resolution data collection strategy and processing for multipolar charge density analysis using XD and MoPro software

  • Expertise in solid state characterization using IR, UV, NMR, MASS, HPLC, DTA, DSC, TGA, XRPD and Hot Stage Microscopy

  • Expertise in aspherical refinement of small molecules and peptides for improved structure model using database transfer (TAAM) approach.


Cryo-EM Core Facility at CeNT UW features:
1. Glacios 200kV cryo-EM microscope equipped with:

  • Schottky field emission source

  • Ceta D 200kV CCD camera

  • Falcon 3EC 200kV direct electron detection camera

  • Phase plate solution for enhanced contrast

  • Autoloader holding up to 12 grids in a cassette

  • Micro-ED package

  • TEM tomography data acquisition software

  • EPU software for single particle analysis experiments

  • EPU-D for micro-ED data collection


    • 2. Vitrobot for automatic vitrification of biological samples

    3. Glow discharge unit (PELCO easiGlow, Ted Pella) for glow discharging TEM grids

    4. Auxillary equipment for micro-ED sample preparation (sonicator, vacuum chamber)

    5. IT infrastructure for data storage & analysis including storage infrastructure, servers, computational clusters, software



    List of recent publications utilising our facility:

    • Tian Y, Liang R, Kumar A, Szwedziak P, Viles JH. 3D-visualization of amyloid-β oligomer interactions with lipid membranes by cryo-electron tomography. Chem Sci. 2021 Mar 31;12(20):6896-6907. doi: 10.1039/d0sc06426b. PMID: 34123318; PMCID: PMC8153238.
    • Alvarez-Malmagro J, Jablonowska E, Nazaruk E, Szwedziak P, Bilewicz R. How do lipid nanocarriers – Cubosomes affect electrochemical properties of DMPC bilayers deposited on gold (111) electrodes? Bioelectrochemistry. 2020 Aug;134:107516. doi: 10.1016/j.bioelechem.2020.107516. Epub 2020 Mar 20. PMID: 32222670.
    • Alvarez-Malmagro J, Matyszewska D, Nazaruk E, Szwedziak P, Bilewicz R. PM-IRRAS Study on the Effect of Phytantriol-Based Cubosomes on DMPC Bilayers as Model Lipid Membranes. Langmuir. 2019 Dec 17;35(50):16650-16660. doi: 10.1021/acs.langmuir.9b02974. Epub 2019 Dec 5. PMID: 31746606.
    • Cytryniak A, Nazaruk E, Bilewicz R, Górzyńska E, Żelechowska-Matysiak K, Walczak R, Mames A, Bilewicz A, Majkowska-Pilip A. Lipidic Cubic-Phase Nanoparticles (Cubosomes) Loaded with Doxorubicin and Labeled with 177Lu as a Potential Tool for Combined Chemo and Internal Radiotherapy for Cancers. Nanomaterials (Basel). 2020 „Nov 16;10(11):2272. doi: 10.3390/nano10112272. PMID: 33207760; PMCID: PMC7696353.”
    • Piszczatowska K, Czerwaty K, Cyran AM, Fiedler M, Ludwig N, Brzost J, Szczepański MJ. The Emerging Role of Small Extracellular Vesicles in Inflammatory Airway Diseases. Diagnostics (Basel). 2021 Feb 2;11(2):222. doi: 10.3390/diagnostics11020222. PMID: 33540806; PMCID: PMC7913078.

    A formal acceptation of the rules and regulations is required prior to granting access to core facilities equipment and staff consultations. All information about them you can find in the form below:

    To reserve your consultation time, please complete the form below: