Equipment

Robotics in the ANU Centre for Therapeutic Discovery in Motion (Credit: Justin Hasell)

The ACTD Facility is comprised of two main screening workstations, the High-Throughput Workstation, and the High-Content Workstation. These workstations are operated independently of each other, are fully automated, and are equipped with various screening equipment, depending on the specific assay requirements.

 

High-Throughput Screening Workstation (HTS)

Fully housed in a Biosafety Class II Cabinet, the HTS workstation contains the following instruments/equipment:

  • 6-axis Denso Robotic Arm and dynamic scheduling control software (Perkin Elmer Plateworks)
  • Barcode Reader
  • Lid Handler (4 lid capacity)
  • Perkin Elmer JANUS Automated Liquid Handling Workstation (enabled for 96/384 tip/pin tool head) and 8-tip Varispan arm (for cherrypicking)
  • Liconic Microplate 37oC CO2 incubator (110 plate capacity)
  • Thermo MultiDrop Combi Liquid Dispenser
  • BioTek ELx405 Plate Washer
  • Plate Hotel (ambient temperature, 42 plate capacity)
  • Brooks Xpeel plate desealer
  • Agilent PlateLoc heated plate sealer
  • Perkin Elmer EnVision microplate reader (details below)
  • Alphascreen Safe Light (Green Light)
  • UV decontamination

 

High-Content Screening Workstation (HCS)

Housed in a Biosafety Class II Cabinet, the HCS workstation contains the following instruments/equipment within and attached to the cabinet:

  • 6-axis Denso Robotic Arm and dynamic scheduling control software (Perkin Elmer Plateworks)
  • Barcode Reader
  • Lid Handler (4 lid capacity)
  • Perkin Elmer Mini JANUS Automated Liquid Handling Workstation (enabled for 96/384 tip/pin tool head)
  • Liconic Microplate 37oC CO2 incubator (110 plate capacity)
  • BioTek ELx406 Plate Washer and Liquid Dispenser
  • Plate Hotel (ambient temperature, 42 plate capacity)
  • Perkin Elmer Opera Phenix High Content Imaging Microscope (details below)
  • IntelliCyt iQue Screener PLUS High-Throughput FACS analyser (details below)

 

Perkin Elmer Opera Phenix High Content Imaging Microscope

Plate/slide types: optimal for 96, 384, 1536 wells (approved plates), glass slides (aqueous mounting media only)

About: The Perkin Elmer Opera Phenix is a sophisticated spinning-disc confocal microscope that contains a microplate stage. While it can be used in the same fashion as a standard confocal microscope, it has been designed for high-throughput, phenotypic screening and assays involving complex disease models, such as live cells, primary cells and micro-tissues. The operating software (Harmony) is also used to develop image analysis algorithms which can take the images that you want to analyse and quantify the specific biology of interest across a large image dataset.

Types of imaging: The imager can be used in either standard (wide-field) or confocal modes, thus allowing imaging of 3D structures, or colocalisation analyses to be easily performed. The compartment where the imaging is performed in the instrument is a sealed chamber, and thus is capable of imaging both fixed cells and live cells. Live cell imaging can be performed at a temperature range of 37-42oC and with CO2 (concentration between 1% - 10%) if required. If there is a requirement for multiple timepoints for imaging, plate manipulation (e.g. addition of reagent) or multiple plates to be imaged in a run, the Phenix can be used in tandem with the ACTD robotics and other components within the HCS cabinet to which the Phenix is attached (liquid dispenser/plate washers/37oC incubator/plate hotel).

Lasers and objectives: The Phenix has 5 excitation lasers and a mixture of air and water objectives, which range from 5x to 63x – this information is provided below. The imager is also capable of capturing label-free brightfield imaging and digital phase imaging. The high NA (Numerical Aperture) water immersion objectives can capture more photons and provides high image resolution even in thick samples. The imager does not have oil-based objectives. Also note that our laser configuration allows FRET-based cellular assays to be performed.

Opera Phenix Objective and Laser/Filter Configuration
Objective Options Excitation Laser Options Emission Filter Options
5x Air (NA 0.16) 375 nm 435-480 nm
10x Air (NA 0.3) 425 nm 435-550 nm
20x Air (NA 0.4) 488 nm 500-550 nm
40x Air (NA 0.6) 561 nm 570-630 nm
20x Water (NA 0.1) 640 nm 650-760 nm
63x Water (NA 1.5)    

 

IntelliCyt iQue Screener PLUS High-Throughput FACS Analyser

Plate/tube types: 96, 384 wells, eppendorf tubes 

About: The IntelliCyt iQue Screener PLUS platform is a high-throughput FACS analyser which is fully integrated with robotics and high-throughput equipment (e.g. liquid dispensing robots, Denso robotic arm) which enables rapid, high content, multiplexed analysis of cells and beads in suspension in both 96 and 384 well plates. The instrument is a voltage-independent system, where altering voltages between plates is not required. Samples are delivered to detectors in an “air gap delimited stream”, and an entire plate of data is processed at one time. As little as 1 μL can be sampled from assay wells. Another feature of the instrument is the enhanced speed of acquisition – minimum sampling time for a 96 and 384 well plate is ~ 5 minutes and ~ 20 minutes, respectively. This makes it highly attractive option for researchers who want to analyse many samples quickly, as well as perform high throughput screens in suspension cells/bead assays. Data is acquired (and can be analysed) using the ForeCyt software, developed specifically to analyse plate-based data rapidly.

Laser Information: The iQue instrument has 3 lasers (Violet-Blue-Red Configuration), and 15 detection channels, with a dynamic range of > 7 decades (non-voltage dependent). The laser/detection configurations are summarised in the table below.

iQue Screener Plus Laser/Detector Configurations
405 nm Laser 488 nm Laser 640 nm Laser
445/45 530/30 675/30
530/30 572/28 780/60
572/28 615/24  
615/24 675/30  
675/30 780/60  
780/60    

 

Perkin Elmer EnVision Microplate Reader

Plate types: 96, 384, 1536 wells 

About: The Perkin Elmer EnVision® microplate reader has capabilities across various measurement technologies including absorbance, fluorescence intensity, fluorescence polarisation (FP), time-resolved fluorescence (TRF), luminescence and AlphaScreen. For automated high-throughput screening, the EnVision is serviced by a Denso Plate Handler 6-axis robot, which is capable of moving plates out of a microplate hotel (at ambient temperature), a temperature-controlled incubator, or from other pieces of equipment into/out of the EnVision instrument. We also have a detachable plate stacker if required.

Technical specifications: 

For absorbance and top-read fluorescence intensity measurements, the instrument features a monochromator that has the following ranges: 

  • Absorbance: 230 – 1000 nm
  • Fluorescence Intensity: 230 – 850 nm 

For filter-based fluorescence intensity, FP, TRF and AlphaScreen measurements we currently have the following filters: 

  • 405/8 
  • 531/25 
  • 535/40 
  • 570/100 (used predominantly for AlphaScreen) 
  • 579/25 
  • 615/5 
  • 642/5 

If the assay requires fluorescence intensity, FP, TRF or AlphaScreen measurements at wavelengths outside of these filter ranges, please discuss this with ACTD staff who will aim to accomodate your needs.