Automated multi-slice extracellular and patch-clamp experiments using the WinLTP data acquisition system with automated perfusion control.

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.

Synaptic plasticity in the hippocampal slice preparation.

Synaptic plasticity is the process by which the brain alters the strength of its synaptic connections, a fundamental function of the brain that enables individuals to learn from experience. The study of synaptic plasticity often involves the application of standard in vitro electrophysiological techniques to hippocampal slice preparations. This unit discusses many of the special considerations that are applicable for the optimal study of synaptic plasticity in this system. Most of these principles also apply to the study of synaptic plasticity in other brain slice preparations.

Capabilities of the WinLTP data acquisition program extending beyond basic LTP experimental functions.

WinLTP is a Windows data acquisition program designed for the investigation of long-term potentiation (LTP), long-term depression (LTD), and synaptic responses in general. The capabilities required for basic LTP and LTD experiments include alternating two-input extracellular pathway stimulation, LTP induction by single train, theta burst, and primed burst stimulation, and LTD induction by low frequency stimulation. WinLTP provides on-line analyses of synaptic waveforms including measurement of slope, peak amplitude, population-spike amplitude, average amplitude, area, rise time, decay time, duration, cell input resistance, and series resistance. WinLTP also has many advanced capabilities that extend beyond basic LTP experimental capabilities: (1) analysis of all the evoked synaptic potentials individually within a sweep, and the analysis of the entire train-evoked synaptic response as a single entity. (2) Multitasking can be used to run a Continuous Acquisition task (saving data to a gap-free Axon Binary File), while concurrently running the Stimulation/Acquisition Sweeps task. (3) Dynamic Protocol Scripting can be used to make more complicated protocols involving nested Loops (with counters), Delays, Sweeps (with various stimulations), and Run functions (which execute a block of functions). Protocol flow can be changed while the experiment is running. WinLTP runs on National Instruments M-Series and Molecular Devices Digidata 132x boards, and is available at www.winltp.com.