Oryx 6
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Oryx Spec
Picture of Oryx
Oryx 6 dispensing routines
Microbatch Crystallization
Getting Started with Oryx
MCC Spec

Oryx 6 for Protein Crystallization with
Microbatch and Sitting Drop

    Oryx 6 Robot       
 

Oryx quickly and accurately dispenses vapor diffusion drops from pre-dispensed reservoirs onto sitting drop shelves in all major high quality vapor diffusion trays available on the market today. Protein is added at the same time using our unique multi-bore dispensing technique.

Our suite of software includes packages for screening and optimization. It provides easy to use interfaces for setting up screening experiments and virtually effortless design of complex two, three and four dimensional multivariate optimization experiments. An extension of the Henderson-Hasselbalch Equation allows linear  interpolation of pH between any two buffer solutions.

Fast, parallel dispensing of all components of optimization experiments deliver up to 384 microbatch wells per hour, automatically covered with oil - with no further requirement for sealing.

 Advantages:

  • Fast

    • Up to 384 completed wells per hour for optimization

    • 96 wells in 12 minutes for sitting drop transfer

    • 96 wells in 50 minutes for microbatch screening experiments

  • Uses little protein

    • Screening experiments as small as 0.1l +0.1l can be dispensed reliably

  • Automatic multivariate designs for optimization
     

  • Affordable
     

  • Microbatch screening finds more leads
     

  • Versatile - offers screening and optimization with a choice of plates and layouts.

Grow large .......     diffractingcrystals    

luciferase.jpg (10193 bytes)
MB crystals of firefly luciferase.  Data could not be collected from VD crystals which were unstable. Courtesy of E. Conti.  Acta Crystallographica. D 52 (1996), 4, pp 876-878
http://www.douglas.co.uk/rep3.htm

 

alcdehydrogenase.jpg (6676 bytes)
MB crystal of alcohol dehydrogenase from a thermophilic bacterium Courtesy of Y. KorkhinActa Crystallographica. D 52 (1996), pp 882-886.
http://www.douglas.co.uk/rep1.htm

   
Crystals of the same alcohol dehydrogenase grown by vapor diffusion in CrystalClear Strips using microseeding. These crystals were used to determine the structure to 2 resolution using synchrotron radiation. Crystallization conditions were determined using the microbatch phase diagram.

 

Software for multivariate experiments
including Central Composite and Box-Behnken (shown) designs
.
    
 box-behn.gif (1411 bytes)
Journal of Crystal Growth 196 (1999) 665 - 673 or
... douglas.co.uk/rat_des.htm

 

Microbatch screening uses very little protein
0.1 - 1 l per well

screen2.gif (2412 bytes)

 

Microbatch optimization is very fast - up to 384 wells/hour

optim2.gif (1195 bytes)

 

Protein crystallization with Crystal Clear strips
sealed with tape
          crystalclrs.gif (3021 bytes)

 

Microbatch screening finds more leads
than VD in a given time
          mbvsvd.gif (3919 bytes)
J. Crystal Growth. 168 (1996), pp 170-174. or: http://www.douglas.co.uk/rep2.htm

 Before reading on, please read the section covering microbatch crystallization, which is essential for a full understanding the operation of Oryx 6.

1. Hardware

Oryx 6 is driven by a PC, which is connected to a stepper motor drive box (MCC). This is connected to 6 motorized syringes (1 large and 5 small) mounted at the back of the chassis, and to the XYZV Plate Loader, which has a moving table and two arms.

 


 
Click here to show full size image

        Small volumes are dispensed by the left-hand arm, using the five small (100 l) motorized syringes.

        Small volumes are dispensed from a two-bore or five-bore microtip.

        Stock solutions do not come into contact with the motorized syringes, which are filled with degassed pure water - this avoids the need for flushing out the syringes when the stock solutions are changed.

        The manual syringes on the front panel are used to refill the motorized syringes, to remove air bubbles, and to load stock solutions.

        Large volumes, including oil for microbatch experiments and reservoir solutions for vapor diffusion experiments, are dispensed by the right-hand arm using the large (10 ml) motorized syringe. A 1 ml disposable tip is used for this.

2. What it does

a. Microbatch screening

The most popular use of Oryx 6 is for screening (finding new crystallization conditions) using the microbatch crystallization method. Here droplets are dispensed into plastic plates and covered with oil to prevent evaporation. (We normally use HLA or Vapor Batch plates. These have wells that hold about 10 μl - we generally dispense 0.2 to 2 l into the bottom of each well.) In order to dispense these very small droplets accurately, a special proprietary microtip is used.

Oryx has an extra liquid handling channel that can be used to cover microbatch droplets with oil immediately after dispensing. This prevents the microtip from coming into contact with the oil, which would significantly reduce accuracy. (This is the reason that Oryx is more accurate than IMPAX for dispensing microbatch experiments.)

Typically 96 premixed solutions can be combined with protein for screening. These solutions are placed on the right-hand side of the Plate Loader. A two-channel microtip is used to mix the solutions with protein. One channel is loaded with the correct amount of protein at the start of the experiment. The other channel is used to pick up 0.1 to 1 l of the first screening solution and transfer it to the first well of the VB plate. It is dispensed and mixed with 0.1 to 1 l of protein from the other channel. Oil is immediately dispensed by the second arm to cover and protect the drop. The microtip is then rinsed. This is repeated for all 96 solutions. It takes about 5 minutes for the user to set up a screening experiment, and 50 minutes to dispense a screen with 96 solutions.

The regular software includes routines for Hampton Research Crystal Screens etc.

Click here for details of the screening liquid handling routines for Oryx.

b. Microbatch optimization

A sophisticated “spreadsheet” environment is provided for designing and carrying out optimization experiments. The stock solutions are shown on the left. Each cell represents one well. The user can input concentrations simply by double-clicking on a well.

By defining the wells at opposite corners of a block, and selecting “interpolate”, the user can rapidly define 2D grids. A similar routine automatically generates a four-dimensional multivariate experimental design. Here the conditions are based on the conditions in a single well – the user chooses the total amount of variation that he would like for each variable as a percentage. See our research report “Practical Experimental Design Techniques” for more details.

The branches of the 5-channel microtip act as reservoirs for protein, precipitant, pH buffer solutions and additives. These are loaded using the lower row of valves and the disposable syringes. Solutions are dispensed using the motorized gas-tight syringes, which contain water.

Exactly the right amount of protein is loaded by sucking it into one bore of the microtip. It is kept separate from the water already in the bore by a small air bubble. Solutions are dispensed from all bores simultaneously. 96 wells can be dispensed in around 15 minutes. As with screening, oil is dispensed by Oryx onto each droplet immediately after it is dispensed.

c. Sitting Drop Crystallization

Oryx can rapidly dispense the droplets into trays such as Crystal Clear, Cryschem, Greiner or Corning plates for sitting drop crystallization. One channel of a 2-bore microtip is loaded with protein at the start of the experiment, then, for each well, reservoir solution is picked up with the other bore and transferred to the droplet. Protein is dispensed at the same time as the reservoir solution and mixed by stirring.

Filling reservoirs for Sitting Drop screens

Oryx has a motorized 10 ml syringe which is connected to a disposable tip. This can be used to transfer solutions from a mother plate to the reservoirs of a sitting drop plate.


Price

Oryx 6 costs 36,000 UK pounds.
We can produce quotations in US$ for US based customers. Due to the current volatility of the US$ the price can no longer be fixed. Please contact us for an up to date quotation.

10% discounts are offered to universities and 5% discounts to government institutes.

Click below for:

Microbatch crystallization

Details of dispensing routines for Oryx 6

Differences between IMPAX and Oryx

Specification of Oryx 6

Specification of MCC control unit

Getting started with Oryx 6

                                                 2004 Douglas Instruments Ltd