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NPI BA-03X Operating Instructions And System Description

Intracellular bridge mode and extracellular amplifier
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OPERATING INSTRUCTIONS AND
SYSTEM DESCRIPTION FOR THE
BA-03X
INTRACELLULAR BRIDGE MODE
and
EXTRACELLULAR
AMPLIFIER
VERSION 2.3
npi 2023
npi electronic GmbH, Bauhofring 16, 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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Summary of Contents for NPI BA-03X

  • Page 1 OPERATING INSTRUCTIONS AND SYSTEM DESCRIPTION FOR THE BA-03X INTRACELLULAR BRIDGE MODE EXTRACELLULAR AMPLIFIER VERSION 2.3 npi 2023 npi electronic GmbH, Bauhofring 16, 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

    3.3. Description of the Rear Panel ..................16 4. Headstage ..........................17 4.1. Headstage Elements ..................... 17 5. Setting up the BA-03X ....................... 18 6. Passive Cell Model ......................19 6.1. Cell Model Description ....................19 6.2. Connections and Operation ..................20 7.

  • Page 3: About This Manual

    BA-03X User Manual _________________________________________________________________________________________________________________ About this Manual This manual should help to setup and use the BA-03X system correctly and to perform reliable experiments. If you are not familiar with the use of instruments for intracellular recording of electrical signals please read the manual completely.

  • Page 4: 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 5: Ba-03X Components

    Other configurations are available, e.g. if the BA-03X system is used only for whole cell patch clamp recordings with suction electrodes the BA-03X system can be delivered with adapted calibrations and a low noise / low bias current headstage (see Optional accessories in chapter 2).

  • Page 6: Description Of The Front Panel

    The BA-03X is also suitable for extracellular measurements and stimulation with glass or metal electrodes. An extended CURRENT RANGE x10 allows electroporation of single cells for non- invasive, juxtacellular filling of cells with dyes or plasmids as well.
  • Page 7 BA-03X User Manual _________________________________________________________________________________________________________________ Figure 1: BA-03X front panel view (the numbers are related to those in the text below) ___________________________________________________________________________ version 2.3 page 7...
  • Page 8 BA-03X User Manual _________________________________________________________________________________________________________________ (1) POWER pressure switch Switch to turn POWER on (switch pushed) or off (switch released). POTENTIAL OUTPUT unit The POTENTIAL OUTPUT unit consists of (2) POTENTIAL OUTPUT GAIN, (34) POTENTIAL OUTPUT FILTER HIGHPASS (Hz), (35) POTENTIAL OUTPUT FROM HEADSTAGE (V), (36) POTENTIAL OUTPUT (mV) and (37) POTENTIAL OUTPUT FILTER LOWPASS (Hz).
  • Page 9 The resistance is measured accurately regardless of the correct setting of other front panel elements such as OFFSET, BRIDGE BALANCE etc. Caution: When the ELECTRODE RESISTANCE button is pressed, the BA-03X automatically applies current pulses of 1 nA to the electrode. Therefore, it should not be used during recordings from cells since this current may stimulate or damage the cell.
  • Page 10 BA-03X User Manual _________________________________________________________________________________________________________________ CURRENT RANGE unit The CURRENT OUTPUT unit consists of (8) CURRENT RANGE LED and (9) CURRENT RANGE switch (8) CURRENT RANGE LED The CURRENT RANGE LED indicates the setting x10 (±120 nA) of switch (9) (LED: ON).
  • Page 11 (mV) start stop time Figure 2: buzz function of the BA-03X (16) OFFSET potentiometer Control to compensate for the electrode OFFSET (ten-turn potentiometer, symmetrical, i.e. 0 mV = 5 on the dial), range: 200 mV (see chapter 7.2). ___________________________________________________________________________ version 2.3...
  • Page 12 BA-03X User Manual _________________________________________________________________________________________________________________ (17) CAPACITY COMP. potentiometer Control for the compensation of the input capacitance (ten turn potentiometer, clockwise, range: 0-30 pF, see chapter 7.3). Caution: This circuit is based on a positive feedback circuit. Overcompensation leads to oscillations which may damage the cell.
  • Page 13 BA-03X User Manual _________________________________________________________________________________________________________________ CURRENT INPUT unit The CURRENT INPUT unit consists of (22) (25) ON / OFF switches, (23) STIMULUS INPUT 1 nA/V connector and (24) STIMULUS INPUT 0.1 nA/V connector. (22) ON / OFF switch Switch to enable (ON) or disable (OFF) STIMULUS INPUT via 1 nA/V connector.
  • Page 14 BA-03X User Manual _________________________________________________________________________________________________________________ (25) ON / OFF switch Switch to enable (ON) or disable (OFF) STIMULUS INPUT via 0.1 nA/V connector. Hint: To avoid interferences always switch to OFF position if the INPUT is not used. gated stimulus unit The gated stimulus unit consists of (26) + / 0 / - switch (STEP SIZE switch), (27) GATE (TTL) connector and (28) STEP SIZE digital potentiometer.
  • Page 15 (39) AUDIO monitor potentiometer Potentiometer for setting the volume of the internal speaker. The AUDIO function of the BA-03X converts the potential output into a tone with a frequency equivalent to the amplitude of the potential. Note: Very small signals of only one or a few mV are probably not well monitored by the AUDIO monitor, especially in extracellular recordings.

  • Page 16: Description Of The Rear Panel

    3.3. Description of the Rear Panel Figure 5: BA-03X rear panel view (the numbers are related to those in the text below) Mains connector Plug for connecting the BA-03X to mains.

  • Page 17: Headstage

    SENSITIVITY switch (+1 V to +7 V, 1V/STEP). 4. Headstage The BA-03X comes with the standard headstage (range: 12 nA, voltage range x1 or 120 nA, voltage range x10) for connecting glass electrodes with high resistances or suction electrodes for whole cell patch clamp recordings with lower resistances via an electrode holder (see Figure A low noise / low bias current headstage (range: 1.2 nA, voltage range x1 or 12 nA, voltage...

  • Page 18: Setting Up The Ba-03X

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

  • Page 19: Passive Cell Model

    Now the BA-03X is ready for an initial check with the cell model. 6. Passive Cell Model The BA-03X can be ordered with a passive cell model as an optional accessory. An active cell model is also available on request (for ref. see Draguhn et al. (1997)).

  • Page 20: Connections And Operation

    BA-03X 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 21 BA-03X User Manual _________________________________________________________________________________________________________________  Connect the BNC jack labeled 10M of the cell model to the BNC connector P of the headstage. b) For simulation of an experiment using a sharp electrode  Connect the BNC jack labeled 50M of the cell model to the BNC connector P at the headstage.

  • Page 22: Test And Tuning Procedures

    BA-03X User Manual _________________________________________________________________________________________________________________ 7. Test and Tuning Procedures Important: The BA-03X should be used only in warmed-up condition, i.e. 30 minutes after turning power on. The following test and tuning procedures are necessary for optimal recordings. It is recommended first to connect a cell model to the amplifier in order to perform some basic adjustments and to get familiar with these procedures.

  • Page 23: Offset Compensation

    The pulses were generated using the automated electrode resistance test circuit of the BA-03X. The upper diagram shows an undercompensated capacitance. In the diagram in the middle the capacitance is slightly overcompensated and in the lower diagram it is well compensated.
  • Page 24 BA-03X User Manual _________________________________________________________________________________________________________________ Figure 9: Tuning of the capacitance compensation using a 100 M resistor ___________________________________________________________________________ version 2.3 page 24...

  • Page 25: Bridge Balance

    BA-03X User Manual _________________________________________________________________________________________________________________ 7.4. Bridge Balance If current is passed through an electrode the occurring voltage deflection (potential drop at R affects the recording of membrane potential. Therefore this deflection must be compensated carefully by means of the BRIDGE BALANCE control. This control is calibrated in M and has two ranges selected by a RANGE switch (#20, Figure 1).

  • Page 26: Electrode Selection

    The recorded signal is amplified with the factor set by switch #2, Figure 1 and filtered using the LOWPASS filter and/or HIGHPASS filter (#34, Figure 1). Since the BA-03X is capable of current injection, the amplifier can be used for extracellular stimulation or electroporation as well. Normally, a current of few nA are used for extracellular stimulation.
  • Page 27 BA-03X User Manual _________________________________________________________________________________________________________________ Figure 10: Tuning of the BRIDGE BALANCE using 100 M resistor ___________________________________________________________________________ version 2.3 page 27...

  • Page 28: Intracellular Sample Experiments

    GND at the headstage.  Make the basic settings (see chapter 5). Again: It is of major importance that the BA-03X systems are used only in warmed-up condition, i.e. 20 to 30 minutes after turning power on.
  • Page 29 BA-03X User Manual _________________________________________________________________________________________________________________ begins to oscillate. Test this by overcompensating the electrode capacitance in several positions of the THRESHOLD potentiometer.  Now the system is preadjusted for measurements. Find a cell!  Approach the desired cell. There are several indications that the electrode is very close to...
  • Page 30 BA-03X User Manual _________________________________________________________________________________________________________________ Figure 13: Artifacts caused by the recording electrode. The measurements were done using a cell model with 100 M membrane resistance, 100 pF membrane capacitance and 100 M electrode resistance. A: C and V not compensated (bridge not balanced)

  • Page 31: Sample Experiment Using A Suction (Patch) Electrode

    BA-03X User Manual _________________________________________________________________________________________________________________ 8.2. Sample Experiment using a Suction (Patch) Electrode If suction (patch) electrodes are used for whole cell recordings they are usually called “pipettes”. Thus, in this subchapter “pipette” means “suction electrode”. Figure 14: Model circuit for whole cell patch clamp recording using a suction electrode...

  • Page 32: Recordings With The Differential Headstage (Optional)

    The drawback is that noise is amplified as well. Therefore, the headstage of the BA-03X can be equipped with a differential input that minimizes noise pick-up. Differential means, that the signal for the amplifier is the difference between the positive (+) (P ) and negative (-) (REF.) input of the headstage.
  • Page 33 BA-03X User Manual _________________________________________________________________________________________________________________ Figure 16: headstage connections, A: differential measurement, B: single-ended measurement Differential measurements can also be done in intracellular recordings. The electrode connected to P is inserted into a cell, and the reference electrode connected to REF is positioned outside the cell.

  • Page 34: Trouble Shooting

    BA-03X User Manual _________________________________________________________________________________________________________________ 9. Trouble Shooting In the following section some common problems, possible reasons and their solutions are described. Important: Please note that the suggestions for solving the problems are only hints and may not work. In a complex setup it is impossible to analyze problems without knowing details. In case of trouble always contact an experienced electrophysiologist in your laboratory if possible, and connect a cell model to see whether the problem occurring with electrodes and “real”...

  • Page 35: Literature

    BA-03X User Manual _________________________________________________________________________________________________________________ The amplifier does not provide any current. Possible reason: 1. The OSCILLATION SHUT OFF circuit is on (LED #30, Figure 1 is red) Solution: 1. Turn the CAPACITY COMP. potentiometer (#17, Figure 1) to the most left position and compensate the input capacitance again.
  • Page 36 BA-03X User Manual _________________________________________________________________________________________________________________  Ogden DC (1994) Microelectrode Techniques. The Plymouth Workshop Handbook, Second Edition, The Company of Biologists Limited, Cambridge  Prinz, A. A. and P. Fromherz (2000). Electrical synapses by guided growth of cultured neurons from the snail Lymnaea stagnalis, Biol. Cybern. 82, L1-L5 ...
  • Page 37 BA-03X User Manual _________________________________________________________________________________________________________________  Rathenberg, J., Nevian, T. & Witzemann, V. (2003). High-efficiency transfection of individual neurons using modified electrophysiology techniques. J Neurosci.Methods. 126, 91-98.  Roberts, W. M., & Almers, W. (1992). Patch Voltage Clamping with Low-Resistance Seals: Loose Patch Clamp.

  • Page 38: Technical Data

    BA-03X User Manual _________________________________________________________________________________________________________________ Technical Data Headstage: Input voltage range: ±1000 mV Operating voltage: ±15 V Enclosure: Size: 23 x 70 x 26 mm, grounded Size: length 150 mm,  8 mm Holding bar: Electrode connector: BNC with driven shield Ground connector: 2.4 mm connector...
  • Page 39 BA-03X User Manual _________________________________________________________________________________________________________________ Outputs: 50  Resistance: Current: BNC connector, sensitivity selectable by rotary switch 0.1, 0.2, 0.5, 1, 2, 5 or 10 V/ nA, display XX.XX nA (CURRENT RANGE: x1), display XXX.X nA (CURRENT RANGE: x10) Potential: BNC connector (POTENTIAL OUTPUT), sensitivity...

  • Page 40: Index

    BA-03X User Manual _________________________________________________________________________________________________________________ Index abbreviations 3 gated stimulus unit 14 accessories 5 general description 5 AUDIO potentiometer 15 GROUND connector 15 basic settings 18 headstage 17 bias current 34 elements 17 bias current adjustment 22 HEADSTAGE connector 12 BIAS current potentiometer 10...

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