Polydimethylsiloxane (“PDMS”)) base material can be cured with deep ultraviolet (DUV) radiation, which in certain examples can obtain a DUV PDMS resolution of about 10 micrometers. See, e.g., K. Tsougeni et al., Microelectronic Engineering 84 (2007) 1104-1008. As seen from
In various embodiments, the properties of a cross-linkable polymer can be changed by modifying the degree of cross-linking. The degree of cross-linking can be modified on a localized basis using lithographic patterns in which the cross-linkable polymer can be selectively and controllably subjected to charged particles or electromagnetic radiation. The modification of the degree of cross-linking can be applied to substrates having surfaces with varying geometric forms. In various embodiments, an elastomer on a substrate is modified to have regions of locally varying rigidity.
This summary is intended to provide an overview of subject matter of the present patent application. It is not intended to provide an exclusive or exhaustive explanation of the invention. The detailed description is included to provide further information about the present patent application.
Embodiments are illustrated by way of example and not limitation in the figures of the accompanying drawings.
In various embodiments, the properties of a cross-linkable polymer can be changed by modifying the degree of cross-linking. The degree of cross-linking can be modified on a localized basis using lithographic patterns in which the cross-linkable polymer can be selectively and controllably subjected to charged particles or electromagnetic radiation. For example, the properties of elastomeric materials, which are cross-linkable polymers, can be changed by modifying the degree of cross-linking of the elastomeric material. The modification of the degree of cross-linking can be applied to substrates having surfaces with varying geometric forms. Such geometric forms on the surfaces can include, but are not limited to, pillars, grooves, ridges, and other forms.
The present inventors have recognized, among other things, that mechanical rigidity of a living or non-living artificial environment can impact motility, adhesion, differentiation, or other behavior or function of living cells. For example, environmental rigidity can play a role in cancerous cell growth. In another example, environmental rigidity can play a role in stem cell differentiation. The present inventors have also recognized that EBL makes it possible to modify the rigidity of structures at the nanoscale, including locally modifying rigidity of a structure at the nanoscale.
In various embodiments, an elastomer is subjected to controlled dosages of charged particles or electromagnetic radiation to selectively adjust a rigidity at localized portions of the elastomer. The elastomer can be disposed on a substrate with a pattern selected based on a criteria for the pattern according to the application in which it is to be implemented. The charged particles can include electrons or ions. The electromagnetic radiation can include UV radiation, DUV radiation, radiation in the visible spectrum, or other range of electromagnetic energy depending on the application for the elastomeric material. The electromagnetic radiation may be used with a photoinitiator for the elastomeric material. The photoinitiator can be in a form that absorbs electromagnetic radiation in applications in which the elastomeric material is transparent to the electromagnetic radiation. The added material absorbs the light such that cross-linking in the elastomeric material is initiated.
In various embodiments, a structure comprises a substrate and a cross-linkable polymeric material disposed on the substrate, where the cross-linkable polymeric material has a surface of locally varying rigidity corresponding to a selected geometric form on the substrate. The cross-linkable polymeric material can include an elastomeric material. The elastomeric material can include a biocompatible polymeric material, such as PDMS. The rigidity of the cross-linkable polymeric material can be selected, but is not limited to, having rigidity values between about 200 Pa and about 1 GPa.
In an illustrative example, an at least partially cured biocompatible polymeric starting material (e.g., silicone rubber or PDMS) is provided. The starting material is selectively exposed to energy, such as an electron-beam provided during electron beam lithography (EBL). This performs selective curing that increases the cross-linking in specified regions, thereby solidifying or otherwise locally increasing the rigidity of such selectively exposed regions relative to other regions that are not so exposed. The selective exposure can be used to generate a specified pattern or structure providing a variable rigidity microenvironment, such as for living cells, a microfluidic application, or the like. A biocompatible material, such as PDMS, that is cross-linkable can be used to create an environment for living cells. An electron discharge layer can be provided before the EBL is carried out.
The present inventors have recognized, among other things, that EBL can be used to selectively and controllably expose the pillars to a current of accelerated electrons. This can change the cross-linking of the polymer in the pillar by a desired degree, thereby increasing its rigidity by a desired degree. By locally altering the electron exposure dosage, regions of different rigidities can be created on the same starting material. Individual pillars can have their rigidity individually adjusted, as desired. Thus, this technique can be used to obtain variable-rigidity surfaces, sub-micron scale rigidity variation, or to determine the interaction between a living cell and an extracellular matrix (ECM).
In various embodiments, a biocompatible polymeric structure, such as PDMS with a patterned surface, has locally varying rigidity corresponding to rigidity values selected to affect a characteristic of a living cell and has a spatial dimension, associated with the rigidity values, that is also selected to affect the characteristic of the living cell. The rigidity values and the dimension of the localized spatial region can be selected to affect cancerous cell growth. The rigidity values and the dimension of the localized spatial region can be selected to affect stem cell differentiation. The biocompatible structure can have varying structural forms with rigidity values that can be selected to be between about 10 kPa and about 1 MPa.
Illustrative examples of an EBL selective rigidity enhancement of PDMS are described below. In an illustrative example, a 20 micrometer thick layer of PDMS can be spun-on or otherwise formed onto a silicon substrate, and then pre-baked for approximately 1 minute at a temperature of approximately 50 degrees Celsius. Supportive substrates are not limited to silicon substrates. EBL can then be performed, such as at an exposure dose of between about 30 μC/cm2 and about 80 μC/cm2. This can be followed by a post-bake for approximately 5 minutes at about 120 degrees Celsius. The PDMS substrate can then be developed, such as by using IPA:MIBK 1:1 for an approximately 3 minute developing time period.
In
The electron beam exposure can be used to obtain varying rigidity of a pre-cured polymeric surface, such as a PDMS surface, which can either be flat, or can have pillars or other surface topography. The below table illustrates various examples of surface type and thickness, surface treatment, whether a degas function is used before exposure to EBL, and the cure time and temperature of the starting PDMS sample.
In an experiment, when cured PDMS was exposed to an electron beam without providing a conducting discharge layer (such as a AuPd sputter layer), blistering and delamination resulted, as illustrated in
In another example, a pre-molded pillared PDMS surface was selectively exposed to EBL.
When modifying an elastomer that has been patterned in micron scaled pillars that are flexible, critical point drying can be used to avoid problems of pillar bending and sticking together of pillars. When the surface of the elastomer having pillars is placed into a liquid, such as water, and removed, the surface tension of the liquid pulls on the pillars and can cause them to distort. For example, Aqua Save applied to an elastomer, such as PDMS, can be removed after the lithography operation in various example embodiments. Removing Aqua Save can involve dipping it in water, where the Aqua Save dissolves. When the structure is removed from the water, water, which has a high surface tension, bends the pillars and sometimes causes pillars to stick to one another. This can be avoided by performing critical point drying. Critical point drying involves taking the structure from water while it is still wet, putting it into another liquid typically alcohol, such as ethanol, and then transferring from the alcohol to carbon dioxide. The carbon dioxide, which starts out in liquefied form, is then placed under high pressure at elevated temperature. It undergoes a phase transformation through its critical point. Critical point drying allows the carbon dioxide to go through the critical point, again at elevated pressure and temperature, basically turning into a gas. There is no evaporation involved such that the carbon dioxide goes immediately from liquid to gas through the critical point without causing a surface tension problem, which avoids the collapse or the sticking of flexible pillars.
The experiments and examples described herein demonstrate, by way of example, but not by way of limitation, that an elastomer, such as a PDMS, can be cured by electron beam exposure. EBL can modulate rigidity of cured PDMS within a readily-accessible dose range. This can help in constructing custom-made substrates for different types of living cells. Such substrates can be used to test cell response to varying rigidity, among other things. In addition or alternative to EBL, UV and DUV light exposure can also be used to selectively vary the rigidity of a PDMS or other polymeric sample. However, EBL can offer a higher resolution potential than DUV, for example. The rigidity of individual pillars, recessed pit sidewalls, or other microstructures can be measured in various ways, such as by using a nanoindenter, lateral force microscopy, or other technique. Because of the ability to spatially control the electron dose on a localized basis, it is possible to create a surface with a rigidity gradient, or other arbitrarily spatially-varying rigidity. In particular, planar or topographical structures of varying rigidity are useful in creating microenvironmental or large-scale cell assays, such as for studying or use with fibroblasts, stem cells, or other living cells.
The structure shown in
The above detailed description includes references to the accompanying drawings, which form a part of the detailed description. The drawings show, by way of illustration, specific embodiments in which the invention can be practiced. These embodiments are also referred to herein as “examples.” Such examples can include elements in addition to those shown and described. However, the present inventors also contemplate examples in which only those elements shown and described are provided. Although various portions of the above description have emphasized EBL treatment of PDMS, the technique can include EBL, ion beam irradiation, or photonic treatment of a suitable polymer, which can be other than PDMS, if desired.
All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference(s) should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls. In this document, the term “or” is used to refer to a nonexclusive or, such that “A or B” includes “A but not B,” “B but not A,” and “A and B,” unless otherwise indicated.
Method examples described herein can be machine or computer-implemented at least in part. Some examples can include a computer-readable medium or machine-readable medium encoded with instructions operable to configure an electronic device to perform methods as described in the above examples. An implementation of such methods can include code, such as microcode, assembly language code, a higher-level language code, or the like. Such code can include computer readable instructions for performing various methods. The code may form portions of computer program products. Further, the code may be tangibly stored on one or more volatile or non-volatile computer-readable media during execution or at other times. These computer-readable media may include, but are not limited to, hard disks, removable magnetic disks, removable optical disks (e.g., compact disks and digital video disks), magnetic cassettes, memory cards or sticks, random access memories (RAMs), read only memories (ROMs), and the like.
The above description is intended to be illustrative, and not restrictive. For example, the above-described examples (or one or more aspects thereof) may be used in combination with each other. Other embodiments can be used, such as by one of ordinary skill in the art upon reviewing the above description. Also, in the above Detailed Description, various features may be grouped together to streamline the disclosure. This should not be interpreted as intending that an unclaimed disclosed feature is essential to any claim. Rather, inventive subject matter may lie in less than all features of a particular disclosed embodiment. Thus, the following claims are hereby incorporated into the Detailed Description, with each claim standing on its own as a separate embodiment.
This application claims priority to U.S. Provisional Application Ser. No. 61/072,717 filed 2 Apr. 2008, entitled “SYSTEMS AND METHODS FOR VARIABLE RIGIDITY SURFACES,” which application is incorporated herein by reference. All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety.
This invention was made with government support under award number PN2 EY 016586 awarded by the NIH Roadmap for Medical Research from the National Institutes of Health. This invention was also made with government support under NSF Award Number CHE-0641523 awarded by the Nanoscale Science and Engineering Initiative of the National Science Foundation. The government has certain rights in this invention. This invention was also made with government support from the New York State Office of Science, Technology, and Academic Research (NYSTAR). The government has certain rights in this invention.
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/US2009/002069 | 4/2/2009 | WO | 00 | 12/21/2010 |
Number | Date | Country | |
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61072717 | Apr 2008 | US |