METHOD OF FORMING A NANOPORE AND RESULTING STRUCTURE
20200088713 ยท 2020-03-19
Inventors
- William J. Durand (Oakland, CA, US)
- Joseph R. JOHNSON (Redwood City, CA, US)
- Roger Quon (Rhinebeck, NY, US)
Cpc classification
B82Y40/00
PERFORMING OPERATIONS; TRANSPORTING
G01N33/48721
PHYSICS
B82Y15/00
PERFORMING OPERATIONS; TRANSPORTING
B82B1/001
PERFORMING OPERATIONS; TRANSPORTING
B82B3/0019
PERFORMING OPERATIONS; TRANSPORTING
International classification
Abstract
Methods are provided for manufacturing well-controlled, solid-state nanopores in close proximity and arrays thereof. In one embodiment, a plurality of wells and one or more channels are formed in a substrate. Each of the wells is adjacent a channel. A portion of a sidewall of each well is exposed. The portion of exposed sidewall is nearest to the adjacent channel. The portion of the exposed sidewall of each well is laterally etched towards the adjacent channel. A nanopore is formed connecting the wells to an adjacent channel.
Claims
1. A method for forming a plurality of nanopores, comprising: depositing a first layer on a substrate; forming a plurality of wells and one or more channels in the first layer and the substrate, each of the plurality of wells being adjacent a channel of the one or more channels; laterally etching a portion of an exposed sidewall to connect the plurality of wells to the adjacent channel; and forming nanopores connecting each of the plurality of wells to the adjacent channel.
2. The method of claim 1, further comprising depositing a second layer on the first layer, the plurality of wells, and the one or more channels to coat each exposed surface prior to exposing the portion of the sidewall of each of the plurality of wells.
3. The method of claim 2, further comprising selectively etching the second layer from the portion of the exposed sidewall prior to laterally etching the portion of the exposed sidewall.
4. The method of claim 3, wherein the second layer is an oxide comprising layer.
5. The method of claim 3, wherein selectively etching the second layer comprises a liquid acidic etch.
6. The method of claim 1, wherein the substrate comprises a crystal structure.
7. The method of claim 6, wherein laterally etching the portion of the exposed sidewall of the plurality of wells comprises a basic wet etch along the crystal structure of the substrate.
8. The method of claim 1, wherein forming the nanopores comprises applying a voltage.
9. A method for forming a plurality of nanopores, comprising: depositing a first layer on a substrate; forming a first well, a second well, and a channel in the first layer and the substrate, the channel being disposed adjacent to the first well and the second well; forming a first tunnel under the first layer, the first tunnel extending between the first well and the channel; forming a second tunnel under the first layer, the second tunnel extending between the second well and the channel; and forming a first nanopore connecting the first tunnel to the channel and a second nanopore connecting the second tunnel to the channel.
10. The method of claim 9, wherein the first nanopore is disposed less than 1 m from the second nanopore.
11. The method of claim 9, wherein the first nanopore is disposed substantially parallel to the second nanopore.
12. The method of claim 9, wherein the first nanopore is disposed at a substantially right angle to the second nanopore.
13. The method of claim 9, further comprising depositing a second layer on the first layer, the first well, the second well, and the channel to coat each exposed surface prior to forming the first tunnel and the second tunnel under the first layer.
14. The method of claim 13, further comprising selectively etching the second layer from a first portion of an exposed sidewall of the first well and a second portion of an exposed sidewall of the second well prior to forming the first tunnel and the second tunnel under the first layer.
15. The method of claim 9, wherein the first tunnel and the second tunnel are formed by a lateral etch.
16. The method of claim 15, wherein the lateral etch comprises a basic wet etch along a crystal structure of the substrate.
17. A device, comprising: a first layer disposed on a substrate; a first well disposed through the first layer within the substrate; a second well disposed through the first layer within the substrate; a channel disposed through the first layer within the substrate adjacent to the first well and the second well; a first laterally etched nanopore coupled to the first well and the channel; and a second laterally etched nanopore coupled to the second well and the channel, the second nanopore being disposed less than 1 m from the first nanopore.
18. The substrate of claim 17, wherein the laterally etched first nanopore is coupled to the first well through a first pyramid shaped tunnel and the laterally etched second nanopore is coupled to the second well through a second pyramid shaped tunnel.
19. The substrate of claim 17, wherein the first well is disposed less than 1000 nm from the second well.
20. The substrate of claim 17, wherein the second nanopore is disposed less than 1000 nm from the first nanopore.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] So that the manner in which the above recited features of the present disclosure can be understood in detail, a more particular description of the disclosure, briefly summarized above, may be had by reference to aspects, some of which are illustrated in the appended drawings. It is to be noted, however, that the appended drawings illustrate only exemplary aspects and are therefore not to be considered limiting of its scope, and may admit to other equally effective aspects.
[0011]
[0012]
[0013]
[0014] To facilitate understanding, identical reference numerals have been used, where possible, to designate identical elements that are common to the figures. It is contemplated that elements and features of one aspect may be beneficially incorporated in other aspects without further recitation.
DETAILED DESCRIPTION
[0015] Methods are provided for manufacturing well-controlled, solid-state nanopores in close proximity and arrays thereof. In one embodiment, a plurality of wells and one or more channels are formed in a substrate. Each of the wells is adjacent a channel. A portion of a sidewall of each well is exposed, the portion of exposed sidewall being nearest to the adjacent channel. The portion of the exposed sidewall of each well is laterally etched towards the adjacent channel. A nanopore is then formed connecting each well to an adjacent channel. Each nanopore can be spaced a distance less than 1 m from adjacent nanopores.
[0016] Methods disclosed herein refer to formation of solid-state nanopores on a semiconductor chip as an example. It is also contemplated that the disclosed methods are useful to form other microfluidic devices and pore-like structures on various materials, including solid-state and biological materials. Methods disclosed herein also refer to formation of pyramid-shaped tunnels as an example; however, other etched features and any combinations thereof are also contemplated. For illustrative purposes, a silicon substrate is described; however, any suitable substrate materials and dielectric materials, such as glass, are also contemplated.
[0017]
[0018] Prior to method 100, a substrate 202 is provided. The substrate 202 is generally any suitable semiconductor substrate, such as a doped or undoped silicon (Si) substrate. The substrate 202 may have thickness between 200 m to 2000 m. In one embodiment, the substrate 202 is Si having a crystal structure including a <100> plane. In operation 110, a first layer 204 is deposited on the substrate 202, as shown in the cross-sectional view of
[0019] In operation 120, a plurality of wells 206A-206B and one or more channels 208 are formed, as shown in
[0020] To form the wells 206A-206B and channel 208 in operation 120, a first photoresist layer 210 is deposited on the first layer 204. A patterning process is then performed to form the wells 206A-206B and channel 208. Generally, the patterning process includes lithographing or patterning the first photoresist layer 210 and etching, for example by reactive ion etching (RIE), the first layer 204 and the substrate 202. The etching may be a directional etch. The first photoresist layer 210 is then removed.
[0021] The wells 206A-206B and channel 208 may be etched to a depth 213 between 10 nm to 2 m. In one embodiment, the wells 206A-206B and channel 208 are etched to have a depth 213 of about 250 nm. The wells 206A-206B may be spaced a distance 212 of between 20 nm to 500 nm away from the channel 208. The channel 208 may have a width 214 of about 1 nm to 200 nm. In one embodiment, the channel 208 may have a width 214 of less than 100 nm. Thus, the first well 206A may be spaced a distance of less than 1000 nm from the second well 206B.
[0022] In operation 130, a second layer 216, such as a material which exhibits a suitable degree of etch selectivity relative to the first layer 204, for example, an oxide layer, is deposited or grown on the first layer 204, the plurality of wells 206A-206B, and the channel 208 to coat each exposed surface of the chip 200, as shown in
[0023] The second layer 216 may be a KOH etch-resistant layer. In at least one implementation, the second layer 216 comprises SiN. The second layer 216 may be base resistant. The second layer 216 generally comprises any suitable dielectric material with an etch rate that is low relative to SiO.sub.2. Examples of suitable materials for the second layer 216 further include, but are not limited to, Al.sub.2O.sub.3, Y.sub.2O.sub.3, and TiO.sub.2. The etch rate of the second layer 216 compared to the etch rate of SiN is generally greater than about 10:1, for example about 100:1, for example about 1,000:1.
[0024] In operation 140, a portion of the sidewall 222 of each of the wells 206A-206B is exposed, as shown in
[0025] To expose the portion of the sidewall 222, a second patterning process is performed. In the second patterning process, a planarization layer 218 is deposited to provide a planar surface for improved photolithography processes. A second photoresist layer 220 is then deposited on the planarization layer 218. A mask may be aligned with the portions of the sidewall 222 to be exposed. The second patterning process includes lithographing or patterning the second photoresist layer 220 and the planarization layer 218. The second patterning process further includes etching, for example by RIE or by a wet etching process, the second photoresist layer 220 and the planarization layer 218 to expose the portion of the sidewall 222 of the wells 206A-206B.
[0026] In operation 150, the second layer 216 is selectively etched from the portions of exposed sidewall 222 of the wells 206A-206B, as shown in
[0027] To remove the second layer 216 from the portions of exposed sidewall 222, a wet etchant is utilized in one embodiment. For example, a fluoride based etchant, such as dilute hydrofluoric acid (DHF), may be used since oxide is selective to fluoride etches. In another embodiment, an isotropic dry etchant is utilized to remove the second layer 216 from the portions of the exposed sidewall 222. For example, the dry etchant may include a fluorine containing vapor or plasma. In one example, the fluorine containing vapor or plasma includes fluorine ions and/or fluorine radicals. The selective etch may remove the second layer 216 while leaving the first layer 204 intact. The second layer 216 may be selectively removed from the portions of exposed sidewall 222 while retaining the second layer 216 on the side surfaces of the wells 206A-206B, as shown in
[0028] In operation 160, the portions of exposed sidewall 222 are laterally etched towards the channel 208. The lateral etchant may comprise a basic liquid chemistry, for example a KOH dip or by exposure to tetramethylammonium hydroxide (TMAH), as shown in
[0029] The lateral etch comprises etching the substrate 202 in a manner parallel to a planar upper surface of the substrate 202. The lateral etch may be an anisotropic etch. Laterally etching the portions of exposed sidewall 222 towards the channel 208 forms tunnels 224 or paths through the substrate 202 under the first layer 204. The tunnels 224 are pyramid or frustum-shaped, and are parallel to a planar upper surface of the first layer 204. The size of the tunnels 224 may vary depending on the size of the portions of exposed sidewall 222. The tunnels 224 may be etched until only a thin film membrane of the second layer 216 remains between the tunnels 224 and the channel 208.
[0030] The lateral etch may be performed for a predetermined amount of time to etch the substrate 202 along the crystal facets or lattice of the crystal structure. The predetermined period of time is generally determined to reduce or eliminate lateral etch relative to the mask opening. In general, the <100> plane of the Si substrate 202 will etch at a rate that corresponds to the temperature of the solution and the concentration of KOH in H.sub.2O. For most scenarios, KOH will etch the <100> plane of Si at a rate of between about 0.4 nm/s and about 20 nm/s. The rate can be accelerated or retarded by cooling or heating the solution. The portions of the exposed sidewalls 222 may be exposed to the etchant for 0.5 to 5 minutes at a temperature of 0 to 100 degrees Celsius. In one embodiment, a 30% weight of aqueous KOH solution is heated to about 40 degrees, and is applied for about 1 minute.
[0031] In operation 170, a plurality of nanopores 226A-226B is formed to connect the tunnels 224 to the channel 208, as shown in
[0032] The nanopores 226A-226B may be formed by applying voltage to induce dielectric breakdown of the thin film membrane of the second layer 216 remaining between the tunnels 224 and the channel 208, resulting in forming well-controlled, localized, and robust nanopores. The nanopores 226A-226B are formed at the tip of the pyramid or frustum-shaped tunnels 224. One or more electrodes 240 may optionally be formed on the chip 200 in order to apply the voltage. The one or more electrodes 240 may be disposed on the second layer 216, within the wells 206A-206B, and within the channel 208. The one or more electrodes 240 may then be removed following the formation of the nanopores 226A-226B. In another embodiment, the chip 200 comprises electrodes configured to apply the voltage. A glass slide 228 may be deposited on and bonded to the second layer 216.
[0033] The applied voltage generally removes at least a portion of the second layer 216 to form the nanopores 226A-226B, for example, by degrading a portion of the second layer 216. The applied voltage generally includes typical voltages above the breakdown voltage of the second layer 216. For example, the breakdown voltage of silicon oxide is generally between about 2 megavolts (MV)/cm and about 6 MV/cm, or between about 200-600 millivolts (mV)/nm of material. In one aspect, the applied voltage is slightly below the breakdown voltage of the second layer 216 and the current is applied for longer to slowly break down the remaining membrane. In another aspect, the applied voltage is above the breakdown voltage of the substrate material such that the nanopores 226A-226B are blasted therethrough. If the nanopores 226A-226B are formed having a larger size than desired, an oxidation process may be performed to reduce the size of the nanopores 226A-226B. For example, the tip of the pyramid or frustum-shaped tunnels 224 may be oxidized to reduce the size of the nanopores 226A-226B. In one embodiment, the second layer 216 is not deposited on or is removed from a portion of the channel 208 disposed between the tunnels 224. In such an embodiment, the nanopores 226A-226B may be formed using the lateral etch of operation 160, and a voltage need not be applied to form the nanopores 226A-226B.
[0034] Forming at least two wells 206A-206B, and subsequently at least two nanopores 226A-226B, allows the nanopores 226A-226B coupled to the wells 206A-206B to be utilized in pairs, or as dual pores, to sequence macromolecules, such as proteins, and/or biological polymers, such as DNA. For example, the chip 200 may be filled with an electrolyte or conductive fluid comprising biological polymers and/or macromolecules. Single strands of DNA or macromolecules may be passed through the nanopore 226A coupled to the first well 206A through the nanopore 226B coupled to the second well 206B to determine properties of or materials attached to the biological polymers and/or macromolecules. The electric properties include an electric signal, which may change based on the size and/or shape of the DNA base pair. The nanopore 226A coupled to the first well 206A may control the collection rate at which biological polymers and/or macromolecules can be attracted to the nanopore 226A, and the nanopore 226B coupled to the second well 206B may control the speed or rate at which biological polymers and/or macromolecules is passed through the nanopore 226B, or vice versa. In another embodiment, both nanopores 226A, 226B influence the speed at which the biological polymers and/or macromolecules is passed therethrough via application of electric fields having different magnitudes. Thus, utilizing dual nanopores allows the dual nanopores to be in fluid communication with one another, resulting in improved signal-to-noise ratios and a higher capturing rate of the biological polymers and/or macromolecules while still maintaining control.
[0035] Because the nanopores 226A-226B have been formed according to methods disclosed herein, the size and position of the nanopores 226A-226B are well controlled. A well-controlled size of the nanopores 226A-226B is generally a diameter suitable for sequencing a sample of a certain size. In one aspect, the size of the nanopores 226A-226B is about 100 nm or less. In one aspect, the nanopores 226A-226B are between about 5 nm by 5 nm and about 50 nm by 50 nm. In one embodiment, the nanopores 226A-226B have a diameter between about 5 nm and 50 nm. In one embodiment, the nanopores 226A-226B are about 20 nm by 20 nm. In another aspect, the size of the nanopores 226A-226B is between about 1.5 nm and about 1.8 nm, such as about 1.6 nm, which is roughly the size of a single strand of DNA. In another aspect, the size of the nanopores 226A-226B is between about 2 nm and about 3 nm, such as about 2.8 nm, which is roughly the size of double-stranded DNA. A well-controlled position of the nanopores 226A-226B is generally any position on the substrate which is suitable for configuration of one or more nanopores. In one embodiment, the nanopores 226A-226B are spaced less than 1 m away from each other, for example less than 100 nm away from each other.
[0036] In one aspect, the chip 200 includes an array of nanopores 226, as shown in
[0037]
[0038] In
[0039] Each channel 308 of the chip 300 may narrow as the channel 308 extends towards the center of the chip 300. The channels 308 may have a width 330 of about 1 m to 20 m. In one embodiment, the channels 308 have a width 330 of about 10 m. The tunnels 324 may have a length 332 extending from one channel 308 to another channel 308 of about 0.1 m to 0.5 m. In one embodiment, the tunnels 324 have a length 332 of about 0.25 m. In another embodiment, the nanopores 326A-326B are spaced less than 1 m away from each other, for example less than 100 nm away from each other. In
[0040] Since the nanopores 326A-326B are disposed at substantially right angles to one another, the distance between the nanopores 326A and 326B does not depend on the width 330 of a channel 308, as the nanopores 326A-326B are not separated by the channel 308. Having wider channels 308 enables the tunnels 324 to be larger as well. Utilizing a chip 300 having closely spaced nanopores 326A-326B and larger tunnels 324 and channels 308 allows for a greater amount of fluid to pass through the channels 308 and tunnels 324, resulting in less electrical resistance being encountered when sequencing biological polymers and/or macromolecules. As such, higher flow rates and enhanced electrical properties may be achieved, and larger biological polymers and/or macromolecules may be sequenced.
[0041] In
[0042] In
[0043] In one embodiment, each of the three pairs of wells 306A-306B has a distinct function for sequencing biological polymers and/or macromolecules, such as providing different fluid and electrical access to the biological polymers and/or macromolecules. For example, after the nanopores 326A-326B have been formed on the chip 300, a sample-containing solution is generally deposited in a first set of wells 306A-306B and a sample-free solution is deposited over a second set of wells 306A-306B.
[0044] The embodiments of
[0045] Benefits of the present disclosure include the ability to quickly form well-controlled nanopores and nanopore arrays having nanopore pairs formed in close proximity. Disclosed methods generally provide nanopores that are well-controlled in size and in position through a thin film membrane. Methods of manufacturing nanopores of well-controlled size provide improved signal-to-noise ratios and higher biological polymers and/or macromolecules capturing rates while maintaining a high level of control. Single strands of biological polymers and/or macromolecules are able to be captured at a higher collection rate and are able to be transmitted through the nanopores at increased speeds, which increases the change in electric current passing through the nanopore. Therefore, utilizing well-controller nanopore pairs provides for improved reading of the DNA sequence.
[0046] While the foregoing is directed to aspects of the present disclosure, other and further aspects of the disclosure may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow.