SGPP Seattle Crystallization Unit Automation in the Seattle Crystallization Unit Larry DeSoto University of Washington.

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Presentation transcript:

SGPP Seattle Crystallization Unit Automation in the Seattle Crystallization Unit Larry DeSoto University of Washington

SGPP Seattle Crystallization Unit Protein Processing in SCU Protein arrives in the SCU Characterization Lori SDS Page Native Gel BCU Concentration Oleksandr Dynamic Light Scattering UV Spectrometer Concentration Limited Proteolysis Optimization

SGPP Seattle Crystallization Unit Robots in Seattle SCU Protein and Reagents Oryx-6 Hydra II Acapella RoboDesign MicroScope II Refined Optimization Initial Optimization or Screen Database (Autoscore and Image Archive) Harvestable Crystals Crystallographer Review

SGPP Seattle Crystallization Unit Douglas Instruments Oryx-6 Primarily for Optimization Screens under oil - ~ 60 minutes Optimization - ~ 15 minutes

SGPP Seattle Crystallization Unit Hydra II Plus One Very easy protocol configuration 96 Channel Dispenser 100 nL to 100 uL Fast ~ 2 minutes for transfer to reservoir, transfer to well and protein dispense ~ 10 minutes total time including wash cycle

SGPP Seattle Crystallization Unit Acapella Capillary Robot

SGPP Seattle Crystallization Unit Acapella Capillary Crystallization Robot High throughput capillary trials Mix and match dispensing of reagent and protein Unique crystallization experiments Standard capillary crystallization experiments

SGPP Seattle Crystallization Unit Acapella Capillary in Glass with no Air Gap D. Meldrum

SGPP Seattle Crystallization Unit Acapella Capillary Trial in Plastic or Glass with Air Gap D. Meldrum

SGPP Seattle Crystallization Unit RoboDesign Microscope II Automated Imaging Flexible Plate Format Standard SBS Plates (any configuration) Douglas Vapor Batch Plates Acapella Microcassettes Approximately 12 minutes per plate (96 wells) Tightly Integrated with Plate Tracking, Scoring and Database Functions

SGPP Seattle Crystallization Unit RoboDesign Microscope II

SGPP Seattle Crystallization Unit RoboDesign Scoring, Image Archive and Database Software Transfer Function CPXO – Automatic Scoring Software CrystalMation – Plate Database Plate Tracking - Plate Barcode Protein Link Image Scheduling Manual Review and Scoring

SGPP Seattle Crystallization Unit RoboDesign CrystalMation – Plate Setup View

SGPP Seattle Crystallization Unit RoboDesign CrystalMation – Plate Inspection View

SGPP Seattle Crystallization Unit RoboDesign CrystalMation – Plate Inspection View

RoboDesign CrystalTrak – Crystallographer’s Notebook or how we got there from here

SGPP Seattle Crystallization Unit CrystalTrak – Crystallographer’s Notebook Protein Tracking Reagent Tracking Drop and Reservoir Design Plate Design Data/Design Export/Import Tracking and Review of Results

SGPP Seattle Crystallization Unit CrystalTrak – Protein View

SGPP Seattle Crystallization Unit CrystalTrak – Reservoir Design

SGPP Seattle Crystallization Unit CrystalTrak – Drop Design

SGPP Seattle Crystallization Unit CrystalTrak – Review View Douglas Vapor Batch

SGPP Seattle Crystallization Unit CrystalTrak – Review View Capillary Cassette (pfal007201wes)

SGPP Seattle Crystallization Unit CrystalTrak – Results View Capillary with pfal007201wes Inner Diameter of the plastic capillary is 190 microns

SGPP Seattle Crystallization Unit The Missing Link – or what do we do with the reservoir and drop designs? Current Procedure Reservoir designs are made by hand Slow Tedious The Future! Our own Matrix Making Robot Tightly coupled with CrystalTrak Fast No Tubes, Pumps or other paraphernalia to complicate life for hard working crystallographers

SGPP Seattle Crystallization Unit RoboDesign Alchemist I Solution Maker

SGPP Seattle Crystallization Unit Acknowledgments SGPP Wim Hol Martin Criminale SCU Lori Andersen Oleksandr Kalyuzhniy Genomation Deirdre Meldrum Mark Holl Shawn McGuire RoboDesign International NIH - NIGMS