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automated atomic absorption spectroscopy

    atomic absorption spectroscopy   links
automated atomic absorption spectroscopy   Process Analysis & Automation were commissioned by Biofocus Ltd. To design and build an automated multi-atomic absorption spectrometer system. Using four Thermo Electron SOLAAR AA spectrometers (with associated Gilson autosamplers), a Hamilton Microlab SWAP robot and a Kendro stacker, a fully automated and reliable workcell was produced. OVERLORD Workstation (now Overlord3) was used control the workcell, with NetLORD (now Overlord3 remoting) used to control the four AA control PCs. We believe that this is the only fully commercially available automated AA system.

categories
dna science
high content screening
high throughput screening
Luminex bead technology
nanopipetting
surface science
atomic absorption spectroscopy
forensic science
elisa assays
automated hplc

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Ion-Channel studies

     
background

The following information is taken from a talk by Dr Adrian Kinkaid of Biofocus Ltd (Cambridge) presented to the European Laboratory Robotics Interest Group at GSK, Stevenage. Biofocus Ltd offer a fully automated service, and details of the service can be found at their web page www.biofocus.com.

This system integrates equipment from Thermo Elemental, Hamilton, Kendro and Metrologic.

paa offer a wide range of standard and user defined automation workcells. For advice on system integration download:

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System Integration

For a copy of the full Biofocus presentation by Adrian Kinkaid presented at ELRIG entitled "Fully Automated High Throughput Ion Channel Screening", download from the PAA downloads page

 
requirements for an automated ion channel analysis system
  • High-throughput
  • Low false-positive rate
  • Low false-negative rate
  • Direct measure of function
  • Good correlation with electrophysiology
  • Reliability
  • Reproducibility
  • Amenable to miniaturization
  • Low cost
  • hERG used as a model channel
 
ion channel screening technologies available (used for hERG)
Fluorescence-based assays
  • Membrane potential-sensitive dyes
Radioligand binding assays - [3H] Dofetilide Automated electrophysiology
  • Automated two-electrode voltage clamp systems
  • Automated whole-cell patch clamp systems
  • Planar patch clamp techniques
Rubidium efflux assays
  • Cerenkov counting of 86 Rb+
  • Atomic absorption spectrometry of 85 Rb+
 
    chosen technique

Thermo AAS

AA instruments

 

Non-radiometric ion flux was chosen as the method of choice because of high throughput relative to other techniques, low cost, direct measurement of channel activity and does not require radiometric reagents.

The Thermo Elemental AAS (right) was selected as the most suitable instrument for the process.

 
    sample preparation method

SWAP loading Autosampler

Robot loading AA autosampler

 
  1. Cells in 96 well plates
  2. Add dilute compound and incubate
  3. Add High K + Buffer and incubate
  4. Transfer supernatant to deep well block or plate
  5. Make up to 1ml or 330ul with 0.1% CsCl Solution
  6. Seal and Store
  7. Read
 
    analysis method - reading the plates

SWAP taking plate from storage for barcode read

Robot taking plate from storage and passed barcode reader

 

The plates are read in batches of four plates.

  • Remove the plate seals by hand and place in Kendro stacker
  • Take each plate in turn
  • Read the barcode
  • Place plate in the next available AA spectrometer Autosampler location
  • Load up 4 plates
  • Start the AA run (each of the 384 wells are sampled, and the Rubidium content determined)
  • Plates are returned the stacker
  • Results are collated with the plate number and sent to the company server for processing
 
more information

Click to view of download documentdownload app note
Ion-Channel studies


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