Whole Cell Path Configuration

Located in the MSB H519 (Rutgers University), the experimental system resting on a TMC vibration isolation table consists of an Olympus BX41 upright microscope designed specifically for the rigorous demands of challenging electrophysiological experiments. The microscope has infrared/DIC optics, epifluorescence illumination with a turret designed for 6 cubes and is coupled to 1/2" Interline CCD camera (380-1200 NM, 570 lines) and a 1000-line, 2-channel video monitor. System runs NIS Elements for image acquisition and control of illumination sequences. The microscope is mounted on a Sutter Instruments motorized stage translator. A Scientifica PatchStar ultra stable, super smooth micromanipulators with X, Y, Z and a smart-sensor- based virtual approach axes is mounted on a Tholabs gantry system. The platform supports a PM-1 Submerged Slice Chamber (Warner Instruments) with perfusion flow is accomplished with a low-electrical noise, two channel peristaltic pump (Watson-Marlow, 400 series) coupled to a 1 channel feed-back temperature control system (TC-324B, Warner Instruments) and Inline Solution Heater (SH-27B, Warne Instruments) for maintaining recorded slices at physiological temperatures.

Recording are obtained using an Axon Multiclamp 700B amplifier (Molecular Devices), a computer- controlled, dual channel, resistive-feedback patch clamp and high-speed current clamp with two CV- 7B low-noise headstages. The amplifier is coupled to a Axon Digidata 1400A (Molecular Devices), a low-noise high-resolution 16-bit data acquisition system with a maximum sampling rate of 250 kHz per channel for electrophysiology recordings connected to an USB 2.0 interface on a Dell Optiplex 780 desktop running pCLAMP 10 (Molecular Devices) Data Acquisition and Analysis Software capable of automated execution of protocols, recording in gap-free and episodic waveform stimulation, online filtering and leak subtraction and online statistical analysis. A 6L Circulating Water-bath (Cole-Parmer) is used for pre heating solutions which are connected to cylinders for constant source of oxygen. Additionally, the system includes a High Voltage, Bipolar Rechargeable Stimulus Isolator (WPI A365R) designed for stimulus delivery in neurophysiological applications and a 2 channel Tektronix oscilloscope.