Crystallization

Crystallization

Please acknowledge the BioX core facility (RRID:SCR_019310) in publications, on posters, or in talks if you use any instruments in the SIP core facility. Please include BioX's RRID (RRID:SCR_019310) and the grant numbers for instruments funded through instrumentation grants in your acknowledgements. This is a requirement from the funding agencies and is crucial for future funding. Find example text on the Acknowledgement Page. 

Phoenix Dropsetter

Image of Phoenix Dropsetter Instrument

 

The Phoenix DropSetter from Art Robbins is a crystallisation dispenser designed for high-throughput screening. It is perfect for the rapid, automated screening of various crystallisation conditions in 96-well plates, using screens from commercial or custom-designed 96 deep-well blocks. The device is equipped with a 96-syringe head for fast and precise dispensing from 96 deep-well blocks and a Decra nano dispenser to deliver the target formulation, such as proteins, DNA, RNA, complexes, or ligands. The Phoenix can set up both sitting and hanging drop experiments and can also pipette seeding experiments.  The 96-syringe head can dispense from 100 nanolitres to 100 microlitres, while the nano dispenser can handle volumes from 100 nanolitres to over 10 microlitres.

Formulatrix Rock Imager

Image of the Dropsetter insturment

 

We have two Formulatrix Rock Imager 54 systems for the automated imaging of crystals in multi-well plates. These imagers combine a temperature-controlled plate hotel with automated crystal imaging at the same temperature. Our two imagers are set to 4°C and 20°C, respectively. Using the integrated Formulatrix software (Rock Maker and Rock Imager), users can automatically capture images of plates using visible light and cross-polarisation. Each well is imaged with 12 images at different focus levels. The PCs running the software and controlling the imagers are equipped with NoMachine, allowing users to control the imaging remotely.

Stereo  Microscopes

Olympus XZ10 Microscope

 

We have several stereo microscopes with cross-polarisation installed in all crystallisation rooms, including the SZX10 Olympus microscope, which is equipped with a Lumen 200 Fluorescence Illumination System. These microscopes enable users to manually screen plates and handle crystals, including “fishing” them into loops for cryo-freezing and mounting at the temperature at which they were grown.

Crystallization Rooms

crystallisation rooms

 

We have three temperature-controlled, walk-in crystallisation rooms set at 4°C, 20°C, and 30°C, each providing shelf space for hundreds of crystallisation trays. Microscopes are installed in all rooms to allow for easy crystal monitoring and fishing without the need for temperature changes or unnecessary transfer of plates between rooms. Additionally, we have crystallisation incubators set at 12°C and 16°C.

 

 

Tools 

 

BioX offers a wide range of tools for manipulating, fishing, freezing, and room-temperature mounting of crystals. The photo shows just a small selection of what is available. The core director will provide a more detailed summary of the tools during training and assist new users in selecting the appropriate tools for their needs.

 

 

Rules Xtal

  • Users should contact Dr Erbse if they are interested in using the X-ray facility and to schedule hands-on training. New users are encouraged to discuss their planned crystallisation experiments with Dr Erbse during the planning phase of their work. This will allow them to benefit from her expertise and ensure that their experiments can be conducted using the tools available in BioX.
  • All users must complete the online "Radiation Safety Training for X-Ray Machine Users"before they can enter the X-ray facility to receive training from Dr.Erbse.
  • Users must receive hands-on training from Dr Erbse before being allowed to use the XtaLAB MM003 independently. This training consists of at least one session and, where possible, uses user samples and lysozyme crystals.
  • After new users have completed all online and hands on training, users will be given access to the Google calendars for the instruments and access to the X-ray facility. Users must sign up for instrument time using the Google calendars.
  • All the instruments in the X-ray core are located in the basement of JSCBB in the C-Wing, room C1B51, on East Campus. Card access is required at all times. Users will only be given access after completing both online and in person trainings.
  • Users are expected to provide all consumables specific to their experiment.
  • Groups must pay for any parts or repairs due to damage by neglect or carelessness. 
  • Cost for repairs/parts/services resulting from normal wear will be split between all user groups.
  • Please Note - Since repairs/service from normal wear might not be necessary every year, depending on use, these costs might come quite some time after the use dates.
  • Users must sign up using the Google calendar for the drop setter. During use, users must sign the paper log at the instrument.
  • Users must stay with the robot at all times while it is going through a protocol.
  • Users must perform the "after run” wash protocol directly after they are done or if they are leaving for more than one hour.
  • Failure to follow these rules will result in loss of the privilege to use the drop setter.
  • Users must sign up using the Google calendar for the imaging system.
  • Please do not schedule automatic imaging. Instead start imaging manually.
  • Check that plates are returned from imaging to their positions in the plate hotel correctly and that no collision occurred.
  • Remove plates no longer needed from the plate hotel.
  • Delete images no longer needed from the computer.