crystal harvesting is finally becoming reality
They said it would never work, but now it is
becoming reality. Sponsored by a NIH STTR grant, a
team of systems
and robotics engineers at
Square One Systems
Design (Jackson, WY) and BR have designed and built the first
semi-autonomous protein crystal harvesting robot.
It performs all steps from crystal looping to cryo-protection,
hyperquenching, and storage in cryo-pucks ready for shipment to the
synchrotron. Additional room temperature mounting and diffraction
characterization of the crystals is in development.
The first demonstration unit has
installed in Spring 2011 at the Joint Center for Structural Genomics
(JCSG) at the Scripps Research Institute in La Jolla, CA, wher
it will be integrated in the
Rigaku CrystalMation HTPX
pipeline. The instrument is avialbale for demonstration at the Joint Center for Structural Genomics
(JCSG). Please contact
Bernhard Rupp or
Marc Deller at JCSG if you want to
visit and schedule a visit and try robotic harvesting.
to the modular design, the system can be exanded to include
tool exchange, which allows for tasks extending beyond the
conventional harvesting of protein crystals. The system is a true
Universal Micromanipulation Robot (UMR). We presented the device
at the first ever dedicated
workshop on automated crystal harvesting at the
National Synchrotron Light Source (NSLS) user's meeting in
Brookhaven, NY, and at the recent NIH PSI Bottleneck meetings
harvesting station - based on the
Robbins Instruments CrysCam - for the budget conscious
Key features of the robotic
Serial robotic arm:
The UMR is based on an anthropomorphic six-axis serial TX-60 robot
arm by Stäubli, the Swiss world leader
in precision robotics. The arm can be equipped witha tool exchanger
providing enormous flexibility and extended operating capabilities.
Optics and lighting:
High quality 0.75-3x (1:4) zoom optics as used on the
proven CrysCam observing
station provide nominal
magnification of 144x exceeding the magnification of most stereo
distance of ~60 mm (>2 inches) allows comfortable sample
access even at maximum resolution. Automated Z-drive (depth) for
auto-focus features. Various custom lighting scenarios incuding
backlighting, optimized for specific plates. 5 Mpix B/W camera
interfaces to image recognition software.
X-Y sample stage:
Custim designed stage accepts 96-well SBS standard plates. Slides
out of cabinet for access, or can be automatically loaded in fully
Digital image recognition:
Crystals are automatically recognized and
characterized by shape and size. The best crystal is automatically
or user-selected, and the robot acquires a harvesting loop of the
appropriate size. It then completes movement of the
harvesting loop to a location immediately adjacent to the crystal.
Once in position, the robot or user executes a pre-programmed capture
routine that is distilled from typical operator-directed capture
routines. After confirmation of successful looping, cryprotection,
hyperquenching, and pin storage are autonomously conducted.
access: A smart new tape punch cuts a small circular hole
instead of removing the entire tape. This procedure not feasible by
hand reduces evaporation losses and extends drop lifetime by about
an order of magnitude. Compare both images of open wells after 15
minutes (tape removed vs. hole punched). A optional, new sealing
station module allows to seal and preserve wells.
A new drip-technique provides generally applicable
cryo-protection by controlled dripping of low-viscosity
perfluoroether from a solenoid valve onto the looped crystal - no,
they do not wash off! Watch the movie (listed below). The
sample is the hyperquenched (Warkentin et al) assuring the fastest
achievable quench rates.
The positional repeatability of the
robot arm below 1 micron allows harvesting of microcrystals of 10 um
size or smaller using the Thorne microloops. They can also be safely
cryo-protected and hyperquenced using the automated protocols
Publications, presentations, and
The first ever robotically harvested protein crystal yields a high resolution trypsin structure with additive-mediated crystal contacts.
J Appl Crystallogr (2007)
Automated robotic harvesting of protein crystals - addressing a critical bottleneck or instrumentation overkill?
of Structural and
Functional Genomics (2007)
Latest presentation given at the
2010 NIH Bottlenecks meeting:
Movie: Overview of robotic harvesting with the UMR (updated version
of production unit soon)
drip-cryoprotection with perfluoroether oil