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    Patch Clamp

    Uploaded by

    Keerthi Kalesan
    The patch-clamp technique allows researchers to study the properties of individual ion channels. It works by (1) sucking a small piece of cell membrane onto a glass micropipette, forming a high-resistance seal. (2) The micropipette can then sense and clamp the voltage inside, to study currents passing through single channels as they open and close in response to voltage changes. There are different patch configurations that allow studying channels in their native environment or isolated from the cell.

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    © All Rights Reserved
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    Patch Clamp

    Uploaded by

    Keerthi Kalesan
    89 views20 pages
    The patch-clamp technique allows researchers to study the properties of individual ion channels. It works by (1) sucking a small piece of cell membrane onto a glass micropipette, forming a high-resistance seal. (2) The micropipette can then sense and clamp the voltage inside, to study currents passing through single channels as they open and close in response to voltage changes. There are different patch configurations that allow studying channels in their native environment or isolated from the cell.

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    The patch-clamp technique allows researchers to study the properties of individual ion channels. It works by (1) sucking a small piece of cell membrane onto a glass micropipette, forming a high-resistance seal. (2) The micropipette can then sense and clamp the voltage inside, to study currents passing through single channels as they open and close in response to voltage changes. There are different patch configurations that allow studying channels in their native environment or isolated from the cell.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    89 views20 pages

    Patch Clamp

    Uploaded by

    Keerthi Kalesan
    The patch-clamp technique allows researchers to study the properties of individual ion channels. It works by (1) sucking a small piece of cell membrane onto a glass micropipette, forming a high-resistance seal. (2) The micropipette can then sense and clamp the voltage inside, to study currents passing through single channels as they open and close in response to voltage changes. There are different patch configurations that allow studying channels in their native environment or isolated from the cell.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    Patch-clamp Technique

    Patch-clamp: the special case of the


    voltage clamp
    (1) Suck a small
    piece of
    membrane onto
    the tip of a glass
    micropipette
    (~ 1 m in
    diameter)

    Cell

    Patch-clamp: the special case of the


    voltage clamp
    (2) Gigaohm-seal

    R > 1 GOhm

    Cell

    Patch-clamp: the special case of the


    voltage clamp
    (3) Sense

    voltage here,
    inside the
    electrode, and
    use voltage
    clamp to keep
    it constant.

    Cell

    Patch-clamp: the special case of the


    voltage clamp
    (3) Sense

    voltage here,
    inside the
    electrode, and
    use voltage
    clamp to keep
    it constant.

    closed
    open

    +
    Cell

    Patch-clamp: the special case of the


    voltage clamp

    closed

    (3) Turn on the aimed


    potential the inside part
    of the pipette and keep
    it constantly by
    applying the voltage
    clamp technique.

    open
    open

    Cell

    Properties of individual voltagedependent sodium channels


    voltage command

    10 msec

    Properties of individual voltagedependent sodium channels


    1. Individual channels are either open
    or closed (no partial openings)

    Properties of individual voltagedependent sodium channels


    1. Individual channels are either open
    or closed (no partial openings)
    2. Each channel opening is only a
    brief event compared to the total
    duration of the whole cell voltagedependent sodium current.

    The macroscopic
    sodium current

    Properties of individual voltagedependent sodium channels


    1. Individual channels are either open
    or closed (no partial openings)
    2. Each channel opening is only a brief
    event compared to the total duration
    of the whole cell voltage-dependent
    sodium current.
    3. Channel opening and closing is
    variable in duration and latency.

    The macroscopic
    sodium current

    Properties of individual voltagedependent sodium channels


    1. The channels are either in open or
    closed state.
    2. The channel openings are short
    events when compared with the
    macroscopic sodium current.
    3. The time duration and latency of the
    channel openings are variable (case
    sensitive). Might happen to not open
    at all.
    4. The open probability of the channels
    resembles with that of the
    macroscopic current.

    Summation of 300 recordings

    The macroscopic
    sodium current

    Properties of individual voltagedependent sodium channels


    1. Individual channels are either open
    or closed (no partial openings)
    2. Each channel opening is only a brief
    event compared to the total duration
    of the whole cell voltage-dependent
    sodium current.
    3. Channel opening and closing is
    variable in duration and latency.
    4. The overall probability of channel
    opening is similar to the total sodium
    current. Look at the sum of the
    currents from 300 trials.
    5. Sometimes an individual channel
    doesnt open even once.

    Summation of 300 recordings

    The macroscopic
    sodium current

    Properties of individual voltagedependent sodium channels


    1. Individual channels are either open
    or closed (no partial openings)
    2. Each channel opening is only a brief
    event compared to the total duration
    of the whole cell voltage-dependent
    sodium current.
    3. Channel opening and closing is
    variable in duration and latency.
    4. The overall probability of channel
    opening is similar to the total sodium
    current. Look at the sum of the
    currents from 300 trials.
    5. Sometimes an individual channel
    doesnt open even once.
    6. Second openings are rare (because
    of inactivation)

    Summation of 300 recordings

    The macroscopic
    sodium current

    Similarly, individual potassium channels,


    calcium channels, and other channels
    can be studied by patch clamping
    1. Individual channels are either open
    or closed (no partial openings).
    Sometimes more than one channel is
    in a patch.
    2. Each channel opening is only a brief
    event compared to the total duration
    of the whole cell current.
    3. Channel opening and closing is
    variable in duration and latency.
    4. The overall probability of channel
    opening is similar to the whole cell
    current
    5. Second openings can happen if
    theres no inactivation.

    Slowly inactivating K
    current channel (Ram &
    Dagan, 1987)

    The configurations of the patch-clamp technique

    OnCell

    Cell-Attached

    The configurations of the patch-clamp


    technique

    OnCell

    Insideout patch

    The configurations of the patch-clamp technique

    OnCell

    Whole
    Cell

    The configurations of the patch-clamp technique

    Whole
    Cell

    The configurations of the patch-clamp technique

    Whole
    Cell

    outsideout patch

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