US 2014/0228857 A1 · Automated Cell Patch Clamping Method and Apparatus

Abstract

In an automated method for in vivo multiple cell patch clamping, cell patch clamping devices are automatically moved into position and targeted to multiple corresponding cells. Cell contact is determined by analyzing the temporal series of measured resistance levels at the clamping devices as they are moved. The difference between successive resistance levels is computed and compared to a threshold, which must be exceeded for a minimum number of computations before neuron contact is assumed. Pneumatic control methods are used to achieve cell-attached or gigaseal formation and subsequent cell break-in, leading to whole-cell patch clamp formation. An automated robotic system automatically performs patch clamping in vivo, automatically detecting cells according to the methodology by analyzing the temporal sequence of electrode impedance changes. By continuously monitoring the patching process and rapidly executing actions triggered by specific measurements, the robot can rapidly find neurons in the living brain and establish recordings.

Claims20

Dependent claims

A method for automated multiple whole-cell patch clamping, comprising the steps of:

in an automated computer processor-controlled apparatus for multiple cell patch clamping,
localizing a plurality of electrodes, each electrode being associated with one of a plurality of cell patch clamping devices, by causing the tips of the electrodes to be moved to an appropriate location for cell hunting;
moving the tips of the plurality of electrodes by a defined amount;
measuring an electrical property at the electrode tips during or after each iteration of the step of moving;
determining whether or not a target cell has been encountered by one or more of the electrodes by constructing a temporal series of the electrical property measurements during or after at least one iteration of the steps of moving and measuring;
iteratively continuing the steps of moving, measuring, and determining until the temporal series of electrical property measurements for one or more of the electrodes, over a threshold number of consecutive iterations, indicates a predefined signature of electrode change that has passed a pre-set cell detection threshold;
pausing the motion of the electrodes;
initiating connection formation for the one or more electrodes that have encountered a target cell;
repeating the steps of iteratively continuing, pausing, and initiating connection until one or more of the plurality of electrodes has encountered a target cell;
and
assessing whether or not connection formation has been achieved at the electrodes.

8 more elements

An apparatus for automated multiple cell patch clamping, comprising:

a cell patch formation apparatus, comprising:
a plurality of cell patch clamping devices, each cell patch clamping device comprising:
an associated electrode;
a 3-axis linear actuator configured for positioning the cell patch clamping device;
a patch amplifier with computer interface;
and
a programmable linear motor configured for moving the cell patch clamping devices up and down in a temporally precise fashion;
and
a computer interface configured for automated closed-loop control of the programmable linear motors based upon a temporal series of electrical property measurements made at the tips of the electrodes.

6 more elements

A method for controlling an automated multiple cell patch clamping device, comprising the steps of:

in a cell patch formation apparatus, comprising:
a plurality of cell patch clamping devices, each having an associated electrode;
a 3-axis linear actuator configured for positioning each cell patch clamping device;
a patch amplifier with computer interface;
a programmable linear motor configured for moving the cell patch clamping devices in a temporally precise fashion;
and
a computer interface configured for closed-loop control of the programmable motor based upon sequences of electrical property measurements made at the tips of the electrodes,
localizing the electrodes by causing the linear motor to move the tip of the electrodes to an appropriate location for cell hunting;
causing the linear motor to iteratively move the tips of the electrodes by a defined amount;
measuring the electrical property at the electrode tips during or after each iteration of the step of moving;
determining whether or not a target cell has been encountered by any of the cell patch clamping devices by constructing a temporal series of the electrical property measurements during or after at least one iteration of the steps of moving and measuring;
iteratively continuing the steps of moving, measuring, and determining until the temporal series of electrical property measurements for one or more of the electrodes, over a threshold number of consecutive iterations, indicates a predefined signature of electrode change that has passed a pre-set cell detection threshold;
causing the linear motor to pause the motion of the electrodes;
initiating connection formation for the one or more electrodes that have encountered a target cell;
assessing whether or not connection formation has been achieved;
if connection formation has been achieved, optionally initiating break-in and formation of at least one whole-cell patch clamp;
and
optionally verifying formation of the whole-cell patch clamps.

15 more elements

Dependencies are inferred from the claim text.

Automated Cell Patch Clamping Method and Apparatus

Life of this patent

Automated Cell Patch Clamping Method and Apparatus

Life of this patent

Abstract

Claims20