Service / Technology Instance

About Macromolecular Crystallisation, Vestec near Prague, Czech Republic

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Crystallization of Proteins and Nucleic Acids core facility provides services focused on obtaining crystals of biomacromolecules and their complexes. We support both manual and robotic crystallization drops setting under various conditions. Remote control of crystal growth in the crystallization hotel is provided using the web interface over Internet.  

Expertise

The macromolecular crystallisation platform enables in-drop dynamic light scattering measurement to check the quality of the protein sample, robotic setup of 96-well crystallisation plates, incubation at selected temperature from a wide range, and automated monitoring of the crystallisation experiments. Experiments can be stored at 4-30°C (or higher). Dedicated rooms with stereomicroscopes for crystal manipulation are available at 20°C, 10°C and 25°C (or higher).

 

Service Availability:

Remote

Physical

Instruments Available:

Preparation of biomolecule crystal samples for the measurement in the diffractometer. Handling of crystals, testing of cryo-conditions, cryo-cooling of crystalline samples.

Crystallisation plate storage and automated crystallisation monitoring enclosure allowing remote access to crystallisation images and data evaluation. Crystallisation drop images are taken using visible light, polarised light and UV.

The hotel is able to store crystallization plates at 10 °C (SBS format, or Lipidic Cubic Phase) and image them according to the pre-set time schedule in visible light, polarized light and UV light. All the images obtained can be viewed, analyzed and evaluated using of the Rock Maker software. The evaluation automatically includes the Marco (Machine Recognition of Crystallization Outcomes), and all the functionalities of the Rock Maker are also available online via the RockMakerWeb.

Nanoliter-volume crystallisation robot with drop dispensing capability, including setting drops for Lipidic Cubic Phase (LCP) experiments. The 8-tip head aspirates and dispenses drops from 10 nL to 1.5 μL. The instrument is able to set up hanging drop, sitting drop, microbatch, in meso LCP drops, additive and seeding experiments. Proportionally-Controlled Active Humidification prevents sample evaporation and increases experiment reproducibility.

The glovebox equipped with a stereomicroscope allows users to manipulate proteins, usually for crystallisation experiments, under an inert atmosphere (usually nitrogen, other inert gases may be used on request).

A multi-channel (96 channels) pipetting robot for the easy set-up of nanodrop crystallisation plates. This crystallisation robot equipment allows the robotic setup of 96-well crystallisation plates, for the screening of crystallisation conditions (and also for “routine crystal production”) - for proteins, nucleic acids, complexes of biological macromolecules.

SONICC is using Second Harmonic Generation (SHG) and Ultraviolet Two-Photon Excited Fluorescence (UV-TPEF) in a completely automated imager to quickly image high throughput crystallization plates and positively identify protein crystals. SONICC will be connected to the current RI1000 crystallisation hotel and will be able to detect even extremely thin crystals, microcrystals <1 μm, and crystals obscured in birefringent LCP.

SpectroLight 600 for measurement of dynamic light scattering in drops (including crystallisation experiments).

Detection of suitability of samples for crystallization or CryoEM grid preparation, together with in situ time lapse monitoring of protein crystallization and detection of nanocrystals for electron diffraction.

Equipment for preparation of samples suitable for electron diffraction and cryoEM experiments. Vitrobot from Thermo Scientific is able to vitrify samples on grids using liquid ethane. Instrumentation like this is crucial for the reproducibility of the procedure during the process of sample optimization.

Instruct Centre

BIOCEV

BIOCEV

Prumyslova 595

25250, Vestec, Prague

Czech Republic

www.biocev.eu

Macromolecular Crystallisation, Vestec near Prague, Czech Republic

Contacts:

Jan Stránský
Jan Stránský

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Jiri Pavlicek
Jiri Pavlicek
BIOCEV - Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University in Vestec
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