Nanopore flow cells and methods of fabrication

Abstract

Nanopore flow cells and methods of manufacturing thereof are provided herein. In one embodiment a method of forming a flow cell includes forming a multilayer stack on a first substrate, e.g., a monocrystalline silicon substrate, before transferring the multilayer stack to a second substrate, e.g., a glass substrate. Here, the multilayer stack features a membrane layer, having a first opening formed therethrough, where the membrane layer is disposed on the first substrate, and a material layer is disposed on the membrane layer. The method further includes patterning the second substrate to form a second opening therein and bonding the patterned surface of the second substrate to a surface of the multilayer stack. The method further includes thinning the first substrate and thinning the second substrate. Here, the second substrate is thinned to where the second opening is disposed therethrough. The method further includes removing the thinned first substrate and at least portions of the material layer to expose opposite surfaces of the membrane layer.

1 . A flow cell device, comprising:
 a glass substrate; and   a membrane layer disposed on the glass substrate, the membrane layer having a nanopore disposed therethrough, and the nanopore is located in a portion of the membrane layer which spans an opening formed through the glass substrate.   
     
     
         2 . The flow cell device of  claim 1 , wherein the glass substrate is formed of fused silica, borosilicate, or a combination thereof. 
     
     
         3 . The flow cell device of  claim 2 , wherein the nanopore has a diameter of about 100 nm or less. 
     
     
         4 . The flow cell device of  claim 3 , wherein a thickness of the membrane layer is less than about 100 nm. 
     
     
         5 . The flow cell device of  claim 1 , wherein the membrane layer is interposed between a first dielectric layer and a second dielectric layer to form a multilayer stack. 
     
     
         6 . The flow cell device of  claim 5 , wherein
 the first and second dielectric layers are formed of silicon oxide, and   the membrane layer is formed of silicon nitride or silicon oxynitride.   
     
     
         7 . The flow cell device of  claim 5 , wherein respective openings formed through each of the first and second dielectric layers expose opposite surfaces of the membrane layer. 
     
     
         8 . The flow cell device of  claim 5 , wherein the second dielectric layer is disposed on the glass substrate, the membrane layer is in contact with the second dielectric layer, and the first dielectric layer is in contact with the membrane layer. 
     
     
         9 . The flow cell device of  claim 8 , wherein the glass substrate is a first substrate and the flow cell device further comprises a second substrate disposed on and in contact with the first dielectric layer, wherein an opening formed through the second substrate is aligned with the opening formed through the first dielectric layer, and the second substrate is formed of fused silica, borosilicate, or a combination thereof. 
     
     
         10 . A nanopore sensor for biopolymer strand sequencing, comprising:
 a flow cell interposed between a first reservoir and a second reservoir, the flow cell comprising a glass substrate and a membrane layer disposed on the glass substrate, the membrane layer having a nanopore formed therethrough, and the nanopore is located in a portion of the membrane layer which spans an opening formed through the glass substrate.   
     
     
         11 . The nanopore sensor of  claim 10 , wherein the glass substrate is formed of fused silica, borosilicate, or a combination thereof. 
     
     
         12 . The nanopore sensor of  claim 10 , wherein the nanopore has a diameter of about 100 nm or less. 
     
     
         13 . The nanopore sensor of  claim 10 , wherein the first and second reservoirs each contain an electrically conductive fluid and a respective electrode that is in communication with a voltage source. 
     
     
         14 . The nanopore sensor of  claim 13 , wherein the voltage source is configured to produce an ionic current flow through the nanopore. 
     
     
         15 . The nanopore sensor of  claim 14 , wherein the membrane layer is interposed between a first dielectric layer and a second dielectric layer to form a multilayer stack. 
     
     
         16 . The nanopore sensor of  claim 15 , wherein the second dielectric layer is disposed on the glass substrate, the membrane layer is disposed on and in contact with the second dielectric layer, and the first dielectric layer is disposed on and in contact with the membrane layer. 
     
     
         17 . A method of sequencing a biopolymer strand using a nanopore sensor, comprising:
 generating a current between a first reservoir and a second reservoir to draw the biopolymer strand through a nanopore of a flow cell interposed between the first reservoir and the second reservoir, wherein
 the flow cell comprises a membrane layer disposed on a glass substrate, 
 the nanopore is formed through a portion of the membrane layer that spans an opening formed through the glass substrate, and 
 the biopolymer strand comprises a plurality of monomer units that sequentially occlude the nanopore as the biopolymer strand is drawn therethrough; and 
   determining, based on changes in the current as the biopolymer strand is drawn through the nanopore, a monomer unit sequence of the biopolymer strand, wherein changes in the current correspond to differences in one or more characteristics of each of the monomer units sequentially occluding the nanopore.   
     
     
         18 . The method of  claim 17 , wherein the glass substrate is formed of fused silica, borosilicate, or a combination thereof. 
     
     
         19 . The method of  claim 17 , wherein
 each of the first and second reservoirs contain an electrically conductive fluid and an electrode,   each of the electrodes is electrically coupled to a voltage source, and   a voltage from the voltage source is used to generate the current.   
     
     
         20 . The method of  claim 17 , wherein
 the membrane layer is interposed between a first dielectric layer and a second dielectric layer to form a multilayer stack, and   respective openings formed through each of the first and second dielectric layers expose opposite surfaces of the membrane layer.