NPI ELC-03XS Operating Instructions And System Description
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NPI ELC-03XS Operating Instructions And System Description

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OPERATING INSTRUCTIONS AND
SYSTEM DESCRIPTION FOR THE
ELC-03XS
UNIVERSAL AMPLIFIER
for EXTRA & INTRACELLULAR RECORDING,
SINGLE CELL STIMULATION and
ELECTROPORATION
VERSION 2.2
npi 2011
npi electronic GmbH, Hauptstrasse 96, D-71732 Tamm, Germany
Phone +49 (0)7141-9730230; Fax: +49 (0)7141-9730240
support@npielectronic.com; http://www.npielectronic.com

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  • Page 1 OPERATING INSTRUCTIONS AND SYSTEM DESCRIPTION FOR THE ELC-03XS UNIVERSAL AMPLIFIER for EXTRA & INTRACELLULAR RECORDING, SINGLE CELL STIMULATION and ELECTROPORATION VERSION 2.2 npi 2011 npi electronic GmbH, Hauptstrasse 96, D-71732 Tamm, Germany Phone +49 (0)7141-9730230; Fax: +49 (0)7141-9730240 support@npielectronic.com; http://www.npielectronic.com...

  • Page 2: Table Of Contents

    Output configuration....................7 Digital displays ......................7 Oscillation shut-off unit....................7 Penetration unit......................7 3.3. Front Panel View of the ELC-03XS Amplifier ............8 3.4. Description of the Front Panel..................9 4. Description of the Rear Panel ..................... 19 Grounding........................21 5.

  • Page 3: Safety Regulations

    (e.g. for diagnosis or treatment of humans), or for any other life-supporting system. npi electronic disclaims any warranties for such purpose. Equipment supplied by npi electronic must be operated only by selected, trained and adequately instructed personnel.

  • Page 4: Introduction

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 2. Introduction “Loose patch” recordings (or “loose seal” recordings [Roberts & Almers, 1992]) are used to record from single excitable cells without damage, i.e. without a direct access to the cell interior. The first recordings were made around 1960 from muscles cells by Alfred Strickholm long time before “tight seal”...
  • Page 5 ELC-03XS User Manual _________________________________________________________________________________________________________________ Today three methods are used to record electrical in vivo or in vitro:  Recordings using patch (suction) electrodes from single neurons  Whole cell patch clamp technique (tight seal recording, intracellular)  Loose patch technique (loose seal recording, extracellular) ...

  • Page 6: Elc-03Xs Amplifier

    (Bridge balance, CC cap comp, offset, holding controls) are calibrated 10-turn potentiometers. The ELC-03XS is capable to record extracellulary DC or AC coupled, to stimulate with current or voltage and to perform non-invasive juxtasomal filling of cells with dyes or DNA.

  • Page 7: Input Configuration

    TTL signals connected to the rear panel (see chapter 4). Output configuration The ELC-03XS amplifier has two output BNCs for POTENTIAL and one output BNC for the CURRENT signal. The POTENTIAL OUTPUT x1 is a pure DC output that monitors the electrode potential directly from the headstage.

  • Page 8: Front Panel View Of The Elc-03Xs Amplifier

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 3.3. Front Panel View of the ELC-03XS Amplifier Figure 1: ELC-03XS front panel view ___________________________________________________________________________ version 2.2 page 8...

  • Page 9: Description Of The Front Panel

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 3.4. Description of the Front Panel In the following description of the front panel elements each element has a number (in bold) that is related to that in Figure 1. The number is followed by the name (in uppercase letters) written on the front panel and the type of the element (in lowercase letters).
  • Page 10 -12000 time Figure 2: BUZZ function of the ELC-03XS (CC Mode) (5) FREQUENCY potentiometer Potentiometer to set the FREQUENCY of pulses within a train in PULSE mode (see also 2). Range: 100 Hz to 2 kHz. (47) AMPLITUDE potentiometer Potentiometer for setting the AMPLITUDE of pulses in PULSE mode (see also 2).
  • Page 11 ELC-03XS User Manual _________________________________________________________________________________________________________________ (6) CURRENT OUTPUT connector BNC connector providing the current output signal; scaling is set by CURRENT OUTPUT SENSITIVITY (V/nA) switch 46, filter is set by CURRENT OUTPUT FILTER LOWPASS (Hz) switch 7. (7) CURRENT OUTPUT FILTER LOWPASS (Hz) switch 16-position rotary switch to select the corner frequency of the LOWPASS FILTER (range: 20 Hz to 20 kHz) for the CURRENT OUTPUT connector (6).
  • Page 12 ELC-03XS User Manual _________________________________________________________________________________________________________________ (12) BRIDGE BALANCE M potentiometer Potentiometer for balancing the BRIDGE circuit; 10 M / turn, range: 0 to 100 M. The BRIDGE must be balanced correctly before using the series resistance compensation in VC mode, because this measure is taken as value for the series resistance compensation (see also 43 and 44).
  • Page 13 ELC-03XS User Manual _________________________________________________________________________________________________________________ (18) SERIES RESISTANCE COMP. (%) potentiometer Potentiometer for setting the compensation for the SERIES RESISTANCE (VC mode) in percent of the resistance value that is set at the BRIDGE BALANCE potentiometer 12. Therefore, it is important to set the BRIDGE BALANCE correctly before using the series resistance compensation (see also 40 and 41).
  • Page 14 ELC-03XS User Manual _________________________________________________________________________________________________________________ (22) THRESHOLD potentiometer Control to set the activation THRESHOLD of the OSCILLATION SHUT-OFF circuit (potentiometer, linear clockwise, range: 0-1200 mV). (23) DISABLED/RESET switch Switch to DISABLE the OSCILLATION SHUT-OFF unit or RESET the circuit. RESET is done if one wants to reset the circuit after previous activation. After resetting the OSCILLATION SHUT-OFF unit is active again.
  • Page 15 ELC-03XS User Manual _________________________________________________________________________________________________________________ CAPACITY COMPENSATION (VC) unit The CAPACITY COMPENSATION (VC) unit consist of (28) TIME CONSTANT (-FAST) potentiometer and of (32) AMPLITUDE (C-FAST) potentiometer. It compensates for the electrode capacity and functions only in VC mode. Tuning is done by application of voltage pulses to the electrode and trying to get the resulting current pulses as fast as possible using potentiometers 28 and 32.
  • Page 16 With an amplifier with switchable headstage (ELC-SWI) the electrode resistance test works also in enhanced (x100) current mode. Note: If the ELC-03XS is equipped with the optional SEAL TEST circuit, the push button is replaced by a three position switch. In the upper position the ELECTRODE RESISTANCE TEST is carried out and in the lower position the SEAL resistance test is accomplished.
  • Page 17 ELC-03XS User Manual _________________________________________________________________________________________________________________ (18) SERIES RESISTANCE COMP. (%) potentiometer Potentiometer for setting the compensation for the SERIES RESISTANCE in percent of the value that is set at the BRIDGE BALANCE potentiometer 12. Therefore, it is important to set the BRIDGE BALANCE correctly before using the series resistance compensation to avoid oscillations (see also 40 and 41).
  • Page 18 Display for the current at the electrode in XX.XX nA, i.e. 10.00 is 10 nA (19.99 nA max.) Note: If the ELC-03XS is equipped with the optional SEAL TEST circuit, the value of the SEAL resistance is also shown at the CURRENT display in XX.XX G (19.99 G max.).

  • Page 19: Description Of The Rear Panel

    Ten-turn potentiometer for the capacity compensation of the electrode in CC mode (ten turn potentiometer, clockwise, range: 0-30 pF). 4. Description of the Rear Panel Figure 3: ELC-03XS rear panel view MONITORING OUTPUT connectors (1) FILTER CURRENT connector BNC connector providing a voltage monitoring the position of the CURRENT FILTER switch (-7 V to +8 V, 1V/STEP).
  • Page 20 BNC connector for remote control of the bridge mode. A TTL HI (+5 V) signal can be connected here to select the bridge mode remotely. (10) OFF connector BNC connector to switch the ELC-03XS in OFF mode remotely with a TTL HI (+5 V) signal. (11) x10 MODE connector BNC connector to switch the ELC-03XS to x10 mode of operation remotely (TTL HI signal).

  • Page 21: Grounding

    ELC-03XS User Manual _________________________________________________________________________________________________________________ (17) Mains connector Plug socket for the mains power-plug. Important: Check line voltage before connecting the ELC amplifier to power. Always use a three-wire line cord and a mains power-plug with a protection contact connected to ground.

  • Page 22: Setting Up The Elc-03Xs

    _________________________________________________________________________________________________________________ 5. Setting up the ELC-03XS The following steps should help you set up the ELC-03XS correctly. Always adhere to the appropriate safety measures (see chapter 1). After unpacking, the ELC-03XS is attached to the setup by assembling the electrical connections.

  • Page 23: Passive Cell Model

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 6. Passive Cell Model The cell model is designed to be used to check the function of the instrument either 1. just after unpacking to see whether the instrument has been damaged during transport or 2. to train personnel in using the instrument or 3.

  • Page 24: Connections And Operation

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 1, 3: connectors for the headstage, 1: electrode resistance: 50 M, 3: electrode resistance: 10 M GND ground connector, to be connected to GND jack of the headstage CELL: switch for cell membrane representing a membrane of either 50 M and 22 pF (CELL 1) or 200 M...
  • Page 25 ELC-03XS User Manual _________________________________________________________________________________________________________________ For a) and b)  Connect GND of the cell model to GND of the headstage. Important: When using a differential headstage (optional) the REF connector at the headstage must not be left open. It must be connected to ground.

  • Page 26: Headstage

    The use of metal electrodes is possible as well. A differential (miniature) headstage (see Optional accessories in chapter 3.1) for measurements in vivo is also available (see also chapter 8.1 and contact npi for details). 7.1. Headstage Elements Figure 7: ELC-03XS headstage ...

  • Page 27: Headstage Bias Current Adjustment

    Caution: It is important that this tuning procedure is performed ONLY after a warm-up period of at least 30 minutes! The ELC-03XS is equipped with a voltage-to-current converter with a very high output impedance which is connected to the recording electrode. The zero current of this unit is tuned with the BIAS current potentiometer (#49, Figure 1).

  • Page 28: Headstage With X1 / X100 Current Range (Option)

    Figure 8: ELC-XS with switchable headstage, front panel view The extended current range can be set at the front panel of the ELC-03XS with an additional switch labeled RANGE x100 or with a TTL signal applied to the x100 INPUT (TTL) BNC connector at the rear panel of the amplifier (see also chapter 4).

  • Page 29: Introduction Into Experiments

    Figure 10: ELC x1 / x100 switchable headstage Important: Even if the ELC-03XS is modified for the x1 / x100 switchable headstage it can be used as a standard amplifier with a standard, i.e. not switchable headstage. In this case the amplifier will recognize that a standard headstage is connected, and the additional current switch at the front panel and the TTL input at the rear panel are automatically disabled.

  • Page 30: Extracellular Voltage Measurement

    ELC-03XS User Manual _________________________________________________________________________________________________________________ Figure 11: headstage connections, A: differential measurement, B: single-ended measurement 8.2. Extracellular Voltage Measurement Extracellular measurements are usually done in the loose-patch configuration or with special metal microelectrodes. Recordings with extracellular metal electrodes is simple. The electrode is advanced into the region where the recordings will be made using a micromanipulator and the signals are filtered and amplified (see chapter 5 in Lalley et al., 1999 for details) as...

  • Page 31: Extracellular Stimulation And Electroporation

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 8.3. Extracellular Stimulation and Electroporation Cells can be stimulated using current or voltage signals. Stimulation with Current  Approach the cell in VC mode and apply square voltage pulses to the electrode.  Contact the cell, establish the loose-patch and disconnect the voltage signal from the COMMAND INPUT :10 mV connector.

  • Page 32: Intracellular Recording

     Apply stimuli to the cell using the STIMULUS INPUT 1 nA/V BNC connector. Voltage Clamp Recording The ELC-03XS can also be used like a whole-cell patch-clamp amplifier.  Approach the cell in VC mode and apply square voltage pulses to the electrode.

  • Page 33: Literature

    ELC-03XS User Manual _________________________________________________________________________________________________________________ 9. Literature General Recording Methods and Voltage Clamp Technique  Dietzel, I. D., Bruns, D., Polder, H. R. and Lux, H. D. (1992). Voltage Clamp Recording, in Kettenmann, H. and R. Grantyn (eds.) Practical Electrophysiological Methods, Wiley- Liss, NY.
  • Page 34 ELC-03XS User Manual _________________________________________________________________________________________________________________  Strickholm, A. (1961). Impedance of a Small Electrically Isolated Area of the Muscle Cell Surface. J Gen.Physiol. 44, 1073-1088. Tracer injection (juxtasomal filling) and extracellular recording using ELC amplifiers  Bruno, R. M. & Sakmann, B. (2006). Cortex is driven by weak but synchronously active thalamocortical synapses.

  • Page 35: Technical Data

    ELC-03XS User Manual _________________________________________________________________________________________________________________ Technical Data Headstage: Input voltage range: ±12 V Operating voltage: ±15 V Enclosure: Size: 23 x 70 x 26 mm, grounded Mounting plate: Size: 70 mm x 50 mm on request length 150 mm,  8 mm...
  • Page 36 ELC-03XS User Manual _________________________________________________________________________________________________________________ Potential LP filter: 4-pole BESSEL filter (other options available) attenuation: -24 dB/octave, corner frequencies (Hz): 20, 50, 100, 200, 300, 500, 700, 1k, 1,3k, 2k, 3k, 5k, 8k, 10k, 13k, 20k Potential HP filter: 1-pole filter, (other options available)

  • Page 37: Index

    ELC-03XS User Manual _________________________________________________________________________________________________________________ Index basic settings 22 Front Panel View 8 bias current adjustment 27 Grounding 21 BRIDGE unit 18 headstage 26 CAPACITY COMPENSATION (VC) unit elements 26 HEADSTAGE connector 9 cell model 23 Intracellular Recording 32 connections and operation 24...

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