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Presented by 
Dilruba Afrin 
Course : GEB-410 
Dept. of Genetic Engineering & 
Biotechnology 
University of Rajshahi
Introduction 
What is patch clamping? 
Why used patch clamp? 
Basic principle 
Types and configurations 
Application 
Limitations
The patch clamp technique is a refinement of the voltage clamp. 
Erwin Neher and Bert Sakmann developed the patch clamp in the 
late 1970s and early 1980s . Neher and Sakmann received the Nobel 
Prize in Physiology or Medicine in 1991 for this work. 
 The method used by Neher, Sakmann & Steinbach in1978 and 
subsequent refinements by Hamill ,Marty, Neher,Sakmann & 
Sigworth in 1981 have led to techniques for high resolution recording 
of current in excised membrane patches in addition to those that 
remain cell-attached.
 Patch clamping is a widely applied electrophysiological technique 
for the study of ion channels; membrane proteins that regulate the 
flow of ions across cellular membranes and therefore influence the 
physiology of all cells. 
 The technique can be applied to a wide variety of cells, but is 
especially useful in the study of excitable cells such as neurons, 
cardiomyocytes ,muscle fibers and pancreatic beta cells. 
 It can also be applied to the study of bacterial ion channels in 
specially prepared giant spheroplasts.
 Provides access to the inside of the cell. 
• Can insert an electrode into the cell 
• Can change the intracellular fluid 
 Creates a seal impermeable to ion flow. 
• High electrical resistance 
 Allows to measure current through ion channels vs. 
voltage,time,temperature,menthol,concentration etc.
 The patch-clamp technique allows the investigation of a 
small set or even single ion channels. It is thus of special 
interest in the research of excitable cells such as neurons, 
cardiomyocytes and muscle fibers. 
1. In principle, thin glass or quartz pipettes with a blunt end are 
sealed onto the membrane (Figure 2). 
2. Suction is applied to aid the development of a high-resistance 
seal in the gigaohm range.
3. This tight seal isolates the membrane patch electrically, which 
means that all ions fluxing the membrane patch flow into the 
pipette and are recorded by a chlorided silver electrode connected 
to a highly sensitive electronic amplifier. A bath electrode is used 
to set the zero level. 
4. To prevent alterations in the membrane potential, a compensating 
current that resembles the current that is flowing through the 
membrane is generated by the amplifier as a negative feedback 
mechanism (Figure 1). 
5. The membrane potential of the cell is measured and compared to 
the command potential. If there are differences between the 
command potential and the measurement, a current will be 
injected.
6. This compensation current will be recorded and allows 
conclusions about the membrane conductance. 
7. The membrane potential can be manipulated 
independently of ionic currents and this allows 
investigation of the current-voltage relationships of 
membrane channels.
Fig.1 : General principle of patch-clamp 
recordings. A glass pipette 
containing electrolyte solution is 
tightly sealed onto the cell 
membrane and thus isolates a 
membrane patch electrically. 
Currents fluxing through the 
channels in this patch hence flow 
into the pipette and can be 
recorded by an electrode that is 
connected to a highly sensitive 
differential amplifier. In the 
voltage-clamp configuration, a 
current is injected into the cell 
via a negative feedback loop to 
compensate changes in 
membrane potential. Recording 
this current allows conclusions 
about the membrane 
conductance.
Fig. 2: A phase contrast image of a patch pipette attached to 
the membrane of a cultured murine hippocampal neuron.
Several variations of the basic technique 
can be applied, depending on what the 
researcher wants to study – 
 Cell-attached or on-cell patch 
 Inside-out patch 
 Whole-cell recording or whole-cell 
patch 
 Outside-out patch 
 Perforated patch 
 Loose patch 
 Automatic patch clamping 
Diagram showing 
configurations of the 
patch clamp technique
 For the evaluation of antiarrhythmics agents. 
 Used for isolated ventricular myocytes from Guinea pigs to study a 
cardio selective inhibition of the ATP sensitive potassium channel. 
 To identify multiple types of calcium channels. 
 To measure the effect of potassium channel openers. 
 Used in molecular biology. 
 Voltage clamp studies on sodium channels. 
 Used to investigate a wide range of electrophysiological cell properties. 
 Measurement of cell membrane conductance.
 Imparting skillful training performance and recording in during 
single channel recording. 
 Cost of process is expensive. 
 Time consuming. 
 Number of samples required is more at times. 
 Chance of membrane distortion.
Thanks to all

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Patch clamp technique principle

  • 1.
  • 2. Presented by Dilruba Afrin Course : GEB-410 Dept. of Genetic Engineering & Biotechnology University of Rajshahi
  • 3. Introduction What is patch clamping? Why used patch clamp? Basic principle Types and configurations Application Limitations
  • 4. The patch clamp technique is a refinement of the voltage clamp. Erwin Neher and Bert Sakmann developed the patch clamp in the late 1970s and early 1980s . Neher and Sakmann received the Nobel Prize in Physiology or Medicine in 1991 for this work.  The method used by Neher, Sakmann & Steinbach in1978 and subsequent refinements by Hamill ,Marty, Neher,Sakmann & Sigworth in 1981 have led to techniques for high resolution recording of current in excised membrane patches in addition to those that remain cell-attached.
  • 5.  Patch clamping is a widely applied electrophysiological technique for the study of ion channels; membrane proteins that regulate the flow of ions across cellular membranes and therefore influence the physiology of all cells.  The technique can be applied to a wide variety of cells, but is especially useful in the study of excitable cells such as neurons, cardiomyocytes ,muscle fibers and pancreatic beta cells.  It can also be applied to the study of bacterial ion channels in specially prepared giant spheroplasts.
  • 6.  Provides access to the inside of the cell. • Can insert an electrode into the cell • Can change the intracellular fluid  Creates a seal impermeable to ion flow. • High electrical resistance  Allows to measure current through ion channels vs. voltage,time,temperature,menthol,concentration etc.
  • 7.  The patch-clamp technique allows the investigation of a small set or even single ion channels. It is thus of special interest in the research of excitable cells such as neurons, cardiomyocytes and muscle fibers. 1. In principle, thin glass or quartz pipettes with a blunt end are sealed onto the membrane (Figure 2). 2. Suction is applied to aid the development of a high-resistance seal in the gigaohm range.
  • 8. 3. This tight seal isolates the membrane patch electrically, which means that all ions fluxing the membrane patch flow into the pipette and are recorded by a chlorided silver electrode connected to a highly sensitive electronic amplifier. A bath electrode is used to set the zero level. 4. To prevent alterations in the membrane potential, a compensating current that resembles the current that is flowing through the membrane is generated by the amplifier as a negative feedback mechanism (Figure 1). 5. The membrane potential of the cell is measured and compared to the command potential. If there are differences between the command potential and the measurement, a current will be injected.
  • 9. 6. This compensation current will be recorded and allows conclusions about the membrane conductance. 7. The membrane potential can be manipulated independently of ionic currents and this allows investigation of the current-voltage relationships of membrane channels.
  • 10. Fig.1 : General principle of patch-clamp recordings. A glass pipette containing electrolyte solution is tightly sealed onto the cell membrane and thus isolates a membrane patch electrically. Currents fluxing through the channels in this patch hence flow into the pipette and can be recorded by an electrode that is connected to a highly sensitive differential amplifier. In the voltage-clamp configuration, a current is injected into the cell via a negative feedback loop to compensate changes in membrane potential. Recording this current allows conclusions about the membrane conductance.
  • 11. Fig. 2: A phase contrast image of a patch pipette attached to the membrane of a cultured murine hippocampal neuron.
  • 12. Several variations of the basic technique can be applied, depending on what the researcher wants to study –  Cell-attached or on-cell patch  Inside-out patch  Whole-cell recording or whole-cell patch  Outside-out patch  Perforated patch  Loose patch  Automatic patch clamping Diagram showing configurations of the patch clamp technique
  • 13.  For the evaluation of antiarrhythmics agents.  Used for isolated ventricular myocytes from Guinea pigs to study a cardio selective inhibition of the ATP sensitive potassium channel.  To identify multiple types of calcium channels.  To measure the effect of potassium channel openers.  Used in molecular biology.  Voltage clamp studies on sodium channels.  Used to investigate a wide range of electrophysiological cell properties.  Measurement of cell membrane conductance.
  • 14.  Imparting skillful training performance and recording in during single channel recording.  Cost of process is expensive.  Time consuming.  Number of samples required is more at times.  Chance of membrane distortion.