Nanopipettingrobot and Crystalisationhotel with UV-/VIS Camera

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 518584441

We apply for a crystallization pipeline composed of a pipetting robot and a crystallization "hotel". The pipetting robot must be able to dispense the reservoirs in the 96-well plates as well as the nanoliter volume drops for the crystallization drops in a highly precise manner. The applied for crystallization hotel is used for the storage of the dispensed 96-well crystallization plates at a defined and stable temperature and is equipped with a UV-/VIS-camera/microscope which allows automated imaging of the plates as well as manually by the user. Both devices use the same software (“Rock Maker”) and use the same database, connecting them to a crystallization pipeline. Robot: The crystallization of proteins starts with the set-up of initial crystallization experiments. Commonly at least 1000 different crystallization drops with a tiny volume (100 - 200 nl) are set up for the first experiments. The drops are so small in volume to use small sample amounts since some proteins cannot be expressed and purified in large quantities (e. g. membrane proteins), or since some compounds (e. g. ligands, substrates, inhibitors etc.) are extremely expensive (for co-crystallization trials). Due to the fact that very small volumes of 100 nl has to be transferred the dispensing performance of the robot has to be extremely precise. This also leads to the need to have a closed system which is equipped with an adjustable humidifier to prevent these tiny drops from drying out. The robot must be able to handle all kinds of the commonly used technics (e. g. "sitting-drop", "hanging drop", "microbatch" und "Lipid Cubic Phase" (LCP)) and also the corresponding reservoirs of these 96-well plates. Hotel: The hotel must be equipped with a cooling system to enable a reliable and adjustable temperature for the storage of the crystallization plates. Furthermore, an integrated UV-/VIS-camera with microscope functionality and suited software is needed in order to ensure an automated imaging under these defined temperature conditions. Additionally, it is essential that this microscope can be use in a non-automatic manner so that the user can analyze the drops with individual settings. For the schedule of the automated imaging (documentation), the management of all data and the implementation of the relevant experimental parameters of each experiment, a database with a suited software is needed. This in turn means that the software of both, the pipetting robot and the storage hotel, have to be compatible with each other as well as with the used database to make this crystallization pipeline possible.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Nano-Pipettierroboter und Kristallisationshotel mit UV-/VIS-Kamera
• Instrumentation Group: 1060 Dilutoren, Pipettiergeräte, Probennehmer
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professor Dr. Sander Smits