title: Automated multi-slice extracellular and patch-clamp experiments using the WinLTP data acquisition system with automated perfusion control.
abstract: WinLTP is a data acquisition program for studying long-term potentiation (LTP) and other aspects of synaptic function. Earlier versions of WinLTP (J. Neurosci. Methods, 162:346-356, 2007) provided automated electrical stimulation and data acquisition capable of running nearly an entire synaptic plasticity experiment, with the primary exception that perfusion solutions had to be changed manually. This automated stimulation and acquisition was done by using 'Sweep', 'Loop' and 'Delay' events to build scripts using the 'Protocol Builder'. However, this did not allow automatic changing of many solutions while running multiple slice experiments, or solution changing when this had to be performed rapidly and with accurate timing during patch-clamp experiments. We report here the addition of automated perfusion control to WinLTP. First, perfusion change between sweeps is enabled by adding the 'Perfuse' event to Protocol Builder scripting and is used in slice experiments. Second, fast perfusion changes during as well as between sweeps is enabled by using the Perfuse event in the protocol scripts to control changes between sweeps, and also by changing digital or analog output during a sweep and is used for single cell single-line perfusion patch-clamp experiments. The addition of stepper control of tube placement allows dual- or triple-line perfusion patch-clamp experiments for up to 48 solutions. The ability to automate perfusion changes and fully integrate them with the already automated stimulation and data acquisition goes a long way toward complete automation of multi-slice extracellularly recorded and single cell patch-clamp experiments.
authors.affiliation: MRC Centre for Synaptic Plasticity, School of Physiology and Pharmacology, University of Bristol, University Walk, Bristol BS8 1TD, UK. w.w.anderson@bristol.ac.uk
authors.forename: William W
authors.initials: WW
authors.lastname: Anderson
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Fitzjohn
authors.forename: Graham L
authors.initials: GL
authors.lastname: Collingridge
mesh_terms: D000818:Animals; D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods*; D045604:Extracellular Fluid / Q000502:physiology; D017774:Long-Term Potentiation / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods; D018408:Patch-Clamp Techniques / Q000295:instrumentation / Q000379:methods; D010477:Perfusion / Q000295:instrumentation / Q000379:methods*; D013223:Statistics as Topic / Q000379:methods*; D013569:Synapses / Q000502:physiology