Dual Pore - Control and Sensor Device

Abstract

Two-pore devices and method for sequencing are described. A two-pore device can include first chamber, a second chamber, and a third chamber, wherein the first chamber is in communication with the second chamber through a first nanopore, and wherein the second chamber is in communication with the third chamber through a second nanopore. The device can also include sensing circuitry for measuring electrical signals associated with a target at a nanopore, and a control circuitry for controlling motion of the target at a nanopore. The device can include and/or switch between sensing and control modes for each of the first nanopore and the second nanopore. Sequencing methods can implement a two-pore device in relation to translocation of a target through one or more nanopores, switching between sensing and control modes as appropriate, and measuring aspects of the target using in sensing modes.

1 . (canceled) 
     
     
         2 . A nanopore device comprising:
 a first chamber, a second chamber, and a third chamber, wherein the first chamber is in communication with the second chamber through a first nanopore, and wherein the second chamber is in communication with the third chamber through a second nanopore;   a sensing circuitry connected to the first nanopore and configured to apply a constant voltage across the first nanopore and to measure a sensing current across the first nanopore for sensing a charged polymer translocating across the first nanopore; and   a control circuitry connected to the second nanopore and configured to apply a dynamic voltage across the second nanopore, the applied dynamic voltage controlling translocation of the charged polymer across the first nanopore and the second nanopore, wherein the applied dynamic voltage is applied using a direct current-biased alternating current signal source.   
     
     
         3 . The nanopore device of  claim 2 , wherein the sensing circuitry comprises a transimpedance amplifier. 
     
     
         4 . The nanopore device of  claim 3 , wherein the transimpedance amplifier is one of a patch clamp or voltage clamp amplifier. 
     
     
         5 . The nanopore device of  claim 2 , wherein the control circuitry comprises a phase lock loop (PLL). 
     
     
         6 . The nanopore device of  claim 5 , wherein the control circuitry is configured to generate an oscillatory voltage output based on feedback from the sensing circuitry. 
     
     
         7 . The nanopore device of  claim 6 , wherein a phase difference between a frequency of the oscillatory voltage output and the frequency of the sensing current is fixed over time. 
     
     
         8 . The nanopore device of  claim 6 , wherein the oscillatory voltage output is provided to a voltage-controlled amplifier (VCA) that applies the dynamic voltage across the second nanopore. 
     
     
         9 . The nanopore device of  claim 2 , wherein the second chamber is electrically coupled as an electrical return path for both the sensing circuitry and the control circuitry of at least one of the first nanopore or the second nanopore. 
     
     
         10 . A nanopore device comprising:
 a first chamber and a second chamber, wherein the first chamber is in communication with the second chamber through a first nanopore and a second nanopore;   a sensing circuitry connected to the first nanopore and configured to apply a constant voltage across the first nanopore and to measure a sensing current across the first nanopore for sensing a charged polymer translocating across the first nanopore;   and   a control circuitry connected to the second nanopore and configured to apply a dynamic voltage across the second nanopore, the applied dynamic voltage controlling translocation of the charged polymer across the first nanopore and the second nanopore, wherein the applied dynamic voltage is applied using a direct current-biased alternating current signal source.   
     
     
         11 . The nanopore device of  claim 10 , wherein the sensing circuitry comprises a transimpedance amplifier. 
     
     
         12 . The nanopore device of  claim 11 , wherein the transimpedance amplifier is one of a patch clamp or voltage clamp amplifier. 
     
     
         13 . The nanopore device of  claim 10 , wherein the control circuitry comprises a phase lock loop (PLL). 
     
     
         14 . The nanopore device of  claim 10 , wherein the device further comprises a first membrane layer that includes the first nanopore, a second membrane layer that includes the second nanopore, and a conductive middle layer between the first membrane and second membrane layer. 
     
     
         15 . A method comprising:
 loading a sample comprising a charged polymer in a first chamber of a nanopore device, wherein the nanopore device comprises the first chamber, a second chamber, a first nanopore, and a second nanopore, wherein the first chamber and the second chamber are in communication through the first nanopore and the second nanopore;   translocating the charged polymer from the first chamber and through the first nanopore by applying a dynamically altered voltage across the second nanopore via a control circuitry connected to the second nanopore;   sensing the charged polymer by applying a constant voltage across the first nanopore via a sensing circuitry connected to the first nanopore and measuring a sensing current across the first nanopore via the sensing circuitry.   
     
     
         16 . The method of  claim 15 , wherein the dynamically altered voltage is determined based on a feedback signal captured by the sensing circuitry connected to the first nanopore. 
     
     
         17 . The method of  claim 16 , wherein the feedback signal is the sensing current measured across the first nanopore which is a measure of movement of the charged polymer across the first nanopore. 
     
     
         18 . The method of  claim 17 , wherein the measure of movement of the charged polymer is a measure of one of a position, a velocity, or an acceleration of the charged polymer. 
     
     
         19 . The method of  claim 16 , wherein the feedback signal is designed in either a frequency domain or a time domain by using one of feedforward or feedback. 
     
     
         20 . The method of  claim 16 , wherein the feedback signal is designed using an estimator and a filter that are designed to estimate molecule-induced changes in the sensing current. 
     
     
         21 . The method of  claim 15 , wherein the dynamically altered voltage is applied with a frequency range between 0.001 Hz and 100 MHz and an amplitude range between 0.001 mV and 10 V.