Applications and references
From primary screening to safety assessment the IonWorks Barracuda system offers the ideal solution
Primary screening of directed libraries
- Speed and cost efficiency that supports screening 100's of thousands of compounds using one compound concentration per well
Rapid lead identification
- 384-well format is ideal for generating large numbers of concentration response curves
- High success rates with IC50 values from 95% or more of wells
Flexibility to optimize leads and perform safety studies
- Software facilitates assay development - alter experimental conditions during runs similar to manual rigs
- Multiple compound additions ideal for optimizing potency and eliminating more toxic compounds
- Supports with-in well assays that generate cumulative response data similar to manual patch clamp methods
Clonal selection
- Single hole recording is an option for identifying and optimizing cell lines
Voltage-gated ion channels
Many voltage-gated ion channels (VGIC) including Na+, K+, Cl-, and Ca2+ channels have been measured on IonWorks® systems. The same VGIC targets measured on the IonWorks HT and Quattro systems can be measured on the IonWorks Barracuda system.
- Deliver complicated voltage protocols to drive ion channels to steady state and to discover state-dependent drug candidates
- Capable of performing within-well assays that generate cumulative response data similar to manual patch clamp methods
- Numerous VGIC targets have been validated including: Na+, K+, Cl-, and Ca2+
Application notes:
Validation of a multiple compound addition assay for hERG ion channel on the IonWorks® Barracuda system
In this study, a multiple compound addition assay for hERG channel was validated by Molecular Devices. Compared to previously reported cross-plate assay, this "within-well" method examines cumulative concentration-response in a similar manner to manual patch clamp, with unparalleled throughput (up to 6000 data points/hour) and comparable pharmacology.
Download the Application Note >
Development of a Nav1.5 Assay for Prolonged Recording on an Automated Electrophysiology System
Poster presented at Biophysics 2013
An assay for screening and confirming blockers of a voltage-gated sodium channel is developed and validated on IonWorks Barracuda. The system enables continuous voltage clamp, sophisticated voltage protocols, flexible recording parameters, and prolonged assay window, for identification of use-dependent compounds in a drug screening setting.
Download the poster >
Validation of IonWorks Barracuda system for hERG voltage ion channel
Data from the direct transfer of a pre-validated hERG assay from the IonWorks Quattro System to the IonWorks Barracuda System. Time and costs were significantly lower on the IonWorks Barracuda system yet comparable results were produced.
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Ligand-gated ion channels
- Simultaneous compound addition and data acquisition allows the measurement of rapidly activating and desensitizing ligand-gated channels
- Numerous LGIC targets have been validated including: ASIC1a, GABA, and nACh
Application notes:
Rapid Solution Exchange for Recording of Fast-Desensitizing Ligand-Gated Ion Channels on the IonWorks Barracuda System
Using Kv1.3 ion channel data proves that the fluid exchange on the IonWorks Barracuda System is fast and complete, making it applicable for analyzing even rapidly desensitizing LGIC targets such as a7-nAChR.
Download the Application Note >
Assay of ASIC1a Channel with the IonWorks Barracuda System
Data produced from continuous recording during ligand addition in 384 parallel recording sites, with nearly 100% success rate in the PPC mode proving IonWorks Barracuda yields unparalleled throughput for LGIC targets.
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References
A small selection of noteworthy papers and articles.
Population patch clamp improves data consistency and success rates in the measurement of ionic currents.
J Biomol Screen 11: 488-496, 2006. Finkel A, Wittel A, Yang N, Handran S, Hughes J, and Costantin J.
http://www.ncbi.nlm.nih.gov/pubmed/16760372?dopt=Citation
Novel 384-well population patch clamp electrophysiology assays for Ca2+-activated K+ channels.
J Biomol Screen 12: 50-60, 2007. John VH, Dale TJ, Hollands EC, Chen MX, Partington L, Downie DL, Meadows HJ, and Trezise DJ.
http://www.ncbi.nlm.nih.gov/pubmed/17092914?dopt=Citation
Population patch clamp electrophysiology: a breakthrough technology for ion channel screening.
Mol Biosyst 3: 714-722, 2007. Dale TJ, Townsend C, Hollands EC, and Trezise DJ.
http://www.ncbi.nlm.nih.gov/pubmed/17882333?dopt=Citation
Designed for accuracy and ease of use
- Individual runs are fully automated
- Easy access to plates and consumables
- Unique cell delivery system minimizes cell usage - < 5 mL/run
- Ultrasonic tip washer with two solution inputs eliminates compound carryover
- CellPettor delivery of cell suspension minimizes on board cell usage when operating with or without optional cell management
Instrument deck: Two boat positions, for extracellular buffer and cells, two compound plates can be used for multiple additions. The wash station cleans both tips and the electrode plate and provides the user with the option for ultrasonic cleaning.
Reliable data - consistent, reproducible results
At the forefront of developing ways to improve assay robustness and data quality Molecular Devices invented Population Patch ClampTM (PPC) technology, a patented, multiple hole-per-well technique that increases chances of finding targets and reduces well-to-well variability.
- PPC substrate has 64 holes per well to enable seal formation and voltage clamp with up to 64 cells simultaneously
PPC experiment of the acid-sensing ion channel ASIC1a showing how PPC generates highly reproducible data with low variability in 384 well-format
Single hole experiment of the acid-sensing ion channel, ASIC1a, shows variability commonly seen using both conventional and single-hole patch-clamping techniques
Proven, cost-effective PatchPlateTM consumables
- Flow-through design enables rapid fluid exchange over the cell
- Constant fluid level maitained for precise aspiration - use compound preparations up to 3x concentration
- User-defined aspirate and dispense speed and height
- Rapid individual offset correction for each of the 384 electrodes
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Automation solutions
- Walk-away operation for up to 6 hours
- Ingrates with user's own robot, stacker or hotel
- Store cells for up to 6 hours
- On-board centrifuge provides fresh cell preps for every run
- Bar code reader for data tracking
- Automation kit includes:
- Plate/consumable landing pad
- Plate handler
- Cell storage vessel
- Cell centrifuge
IonWorks Barracuda® system
