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The Topaz System
The TOPAZ system uses nanoscale free interface diffusion (FID) to set the new standard for efficiency in protein crystallization. The family of chips includes the 1.96 Diffraction Capable (DC) chip–which uniquely provides early access to high quality in situ X-ray diffraction data–and the 4.96 screening chip–which provides the ideal solution for ultra low-volume, parallel screening.

The TOPAZ system for protein crystallography provides a complete screen-to-beam solution.
  • High quality in situ diffraction-based screening of crystals
  • The lowest sample volume requirements
  • The most accurate crystal calling software in the industry
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TOPAZ® System
Overview

The TOPAZ system for protein crystallography provides a complete screen-to-beam solution.

  • High quality in situ diffraction-based screening of crystals
  • The lowest sample volume requirements
  • The most accurate crystal calling software in the industry

The TOPAZ system sets new standards for efficiency in protein crystallization. It consists of IFCs, instrumentation, and software that simplify and automate nanoscale free interface diffusion (FID). The family of IFCs includes the 1.96 Diffraction Capable (DC) chip–which uniquely provides early access to high quality in situ X-ray diffraction data–and the 4.96 screening chip–which provides the ideal solution for ultra low volume, parallel screening.

Instruments

The TOPAZ system minimizes the manual work associated with protein crystallization. Because the instrumentation and software were co-developed with the TOPAZ chips, all components work synergistically to deliver reliable, reproducible results using nanoscale free interface diffusion (FID).

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TOPAZ IFC Controller

The TOPAZ IFC Controller provides the most efficient and economic solution for setting up FID crystallization screening. Key advantages include:

  • Automated setup and control of FID for Topaz chips
  • Self-contained and fully integrated design
  • Touch screen interface

Researchers simply transfer 96 reagents and samples into wells on the frames of chips and place the chips into the bay of the controller. The IFC Controller does the rest, pressure loading the fluids and tracking experiments. The TOPAZ IFC Controller is designed accept one chip at a time.

topaz2.jpgTOPAZ IFC Controller

AutoInspeX II Workstation for the TOPAZ System

The AutoInspeX II workstation (AIX II) works in synergy with TOPAZ chips to redefine efficiency in automated crystallization analysis. The benefits of the AIX II include:

  • Automated imaging of TOPAZ chips
  • High quality images that are always in focus
  • The most accurate crystal calling capability in the industry

The workstation produces images that are consistently clear and aligned, unlike drop-based platforms, which suffer from variable size, shape and location of drops under the microscope. In addition, Fluidigm now offers an upgrade that equips the workstation for imaging of crystallization experiments in qualified microwell plates.




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AutoInspex II Workstation
Chips

TOPAZ chips provide the most efficient screen-to-beam solution for protein crystallization. Here are a just a few of the advantages of TOPAZ chips:

  • Free Interface Diffusion (FID) and dehydration based experiments
  • Ultra low sample volume requirements
  • Simple, easy to use workflow for setting up experiments

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TOPAZ 1.96 Diffraction Capable Chip

TOPAZ 1.96 Diffraction Capable (DC) chip uses the latest advances in microfluidic technology to enable in situ data collection:

  • In situ diffraction-based screening of crystals
  • Data collection from the entire chip or chip cutouts
  • No handling of individual crystals

High quality X-ray diffraction data can now be obtained early in the screening process, enabling critical decisions to be based on data rather than just subjective assessments, such as visual quality of crystals. The 1.96 DC chip can be used to screen one sample against 96 conditions at two different mixing ratios.

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TOPAZ 1.96 Diffraction Capable Chip

TOPAZ 4.96 Screening Chip

The TOPAZ 4.96 screening chip is the ideal solution for ultra low volume, parallel screening:

  • Only one µl of each sample to screen against 96 conditions
  • Screen up to four samples in parallel
  • Ideal configuration for high throughput screening

The chip requires only one microliter of protein solution per 96-reagent screen.  It also provides the ability to pursue a small volume parallel screening approach to crystallization. The 4.96 screening chip can be used to screen up to 4 samples against 96 conditions each.

After initial crystallization conditions are identified, scaling up to diffraction-quality crystals by using vapor diffusion is made easy by following Fluidigm's recommended translation protocols.
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TOPAZ 4.96 Screening Chip


Software

AutoInspeX II Software Suite: Crystal Vision Software

The TOPAZ system includes the most advanced protein crystallization software analysis tools in the industry. Its advantages include:

  • Best in class crystal scoring capability
  • Images that are always in focus
  • Simple, intuitive interface

Fluidigm's Crystal Vision Software dramatically improves the efficiency of crystal analysis and scoring. The image processing software uses high-resolution data to accurately score the degree of crystallization activity and, thereby, removes much of the subjective analysis burden typically placed on researchers. An entire series of experiments can be evaluated in one session, saving hours of inspection time.

Crystal Vision Software at a Glance:
Calling accuracy: < 1% false negatives
 < 5% false positives
Classification of crystals 10 x 20 micron and larger





Activity scores for the entire 96-reagent screen can be viewed in various tabular formats (Figure 1) or as color-coded layouts of the experiments (Figure 2). This global view is helpful to rapidly determine the most promising conditions for crystallization. Images for individual chambers can be magnified and viewed side-by-side to compare changes over several time points.

If you wish to schedule a software demo, please contact your local sales representative

Crystal Vision Software Screenshots

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Fig. 1. The Table View shows both Auto Calls, "Crystal" or "Ranked" according to activity score, and User Annotations. Data is displayed in an easy-to-use format that speeds comparative analyses.		 cv_layout.jpg
Fig. 2. The Layout View allows quick assessment of crystallization activity across the entire chip run. Blue squares indicate chambers containing crystals. Viewing historical data allows easy comparison of results over several time points.

TOPAZ® Database Suite: Experiment Manager Software

The TOPAZ database suite is used to manage information from every component of the TOPAZ system through a single user-friendly interface. The advantages of the TOPAZ database include:

  • Complete data management from setup to follow on experiments
  • Easy analysis of results across the entire database
  • Comprehensive web-based reports

Experiment Manager Software provides master scheduling and analysis tools to design and execute experiments from initial screening through scale-up. Image data can be sorted and/or queried by automatic or user–annotated results. The data may be compared using these queries or by referring directly to historical image data. Intuitive design wizards and templates make it easy to design follow–on experiments applicable to any crystallization method.

If you wish to schedule a software demo, please contact your local sales representative.

Experiment Manager Software Screenshots

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Fig. 1.
Experiment Manager software automatically integrates OptiMix or third-party reagent information		 EM_design.jpg
Fig. 2.
Experiment Manager software helps you design follow-on experiments.
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Fig. 3.
Experiment Manager software helps you to discover crystallization trends quickly using multiple parameter queries.		 dbxreport.jpg
Fig. 4.
Database Web Reports are customized HTML reports of top crystallization activity accessible from any desktop.
Literature

Instrument Data Sheets

Chip Data Sheets

Application Notes

Other Documentation

  • Research Report - The TOPAZ System Enables Rapid Discovery at San Diego Biotech Company, Rx3 Pharmaceuticals 
  • White Paper - The Advantages of the TOPAZ Approach to Protein Crystallization: A Theoretical Proposition
  • TOPAZ Bibliography - Breakthroughs Enabled by The TOPAZ System for Protein Crystallization

Featured Publications

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Featured Posters

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Workflow/Specs

Workflow


 1. Prep.
Prep the IFC in the TOPAZ IFC controller.  The IFC controller will automatically close the interface valves, preventing premature mixing of samples and reagents.
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2. Transfer.
Pipette the samples and reagents into the appropriate wells on the frame of the chip.
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3. Load.
Load the samples and reagents into the chip by placing it in the TOPAZ IFC controller and selecting the "load" button on the instrument's integrated touchscreen interface.
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4. Image.
Image the chips automatically using the AutoInspeX II workstation according to the recommended schedule.

5. Analyze.
Analyze the results using Crystal Vision™ Software and the TOPAZ database, which provide the most accurate crystal scoring in the industry and the ability to investigate results across experiments.

Specifications

Crystal Vision Software at a Glance:
Calling accuracy: < 1% false negatives
 < 5% false positives
Classification of crystals 10 x 20 micron and larger





TOPAZ IFC Controller at a Glance:
Number of Chips: 1 (Single Bay)
Experiment Tracking: Barcode
Gas Pressure: Internal Compressor
Interface: On-Board Touchscreen





AutoInspeX II Workstation at a Glance:
Image resolution 2600 x 2060 with 0.5 microns/pixel at 10X
Class. threshold:
 10 x 20 micron and larger
Processing time:
 < 4 mins per 1.96 chip imaging
< 14 mins per 4.96 chip imaging
Temp. range:
 4ºC - 35ºC
*Accuracy:
 < 1% false negatives
< 5% false positives





*As demonstrated by the Fluidigm Install Kit