Abstract
The challenge of constructing surfaces with nanostructured chemical functionality is central to many areas of biology and biotechnology. This protocol describes the steps required for performing molecular printing using polymer pen lithography (PPL), a cantilever-free scanning probe-based technique that can generate sub-100-nm molecular features in a massively parallel fashion. To illustrate how such molecular printing can be used for a variety of biologically relevant applications, we detail the fabrication of the lithographic apparatus and the deposition of two materials, an alkanethiol and a polymer onto a gold and silicon surface, respectively, and show how the present approach can be used to generate nanostructures composed of proteins and metals. Finally, we describe how PPL enables researchers to easily create combinatorial arrays of nanostructures, a powerful approach for high-throughput screening. A typical protocol for fabricating PPL arrays and printing with the arrays takes 48–72 h to complete, including two overnight waiting steps.
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Acknowledgements
This material is based on the work supported by the US Defense Advanced Research Projects Agency (DARPA) Microsystems Technology Office (MTO) award N66001-08-1-2044; Asian Office of Aerospace Research and Development (AOARD) award FA2386-10-1-4065; Air Force Office of Scientific Research (AFOSR) awards FA9550-12-1-0280 and FA9550-12-1-0141; US National Science Foundation awards DBI-1152139, DBI-1152169 and DMB-1124131; Department of Defense (DoD)/Naval Postgraduate School (NPS)/National Security Science and Engineering Faculty (NSSEF) fellowship awards N00244-09-1-0012 and N00244-09-1-0071; Chicago Biomedical Consortium with support from Searle Funds at The Chicago Community Trust; and Center of Cancer Nanotechnology Excellence (CCNE) initiative of the US National Institutes of Health (NIH) award U54 CA151880. D.J.E. is supported by a DoD, Air Force Office of Scientific Research, National Defense Science and Engineering Graduate Fellowship (NDSEG) 32 CFR 168a. X.L. gratefully acknowledges support from the Ryan Fellowship from Northwestern University. K.A.B. gratefully acknowledges support from Northwestern University's International Institute for Nanotechnology. B.R. acknowledges the Indo-US Science and Technology Forum (IUSSTF) for a postdoctoral fellowship. L.R.G. acknowledges the NSF for a Postdoctoral Research Fellowship in Biology.
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D.J.E., X.L., K.A.B., L.R.G., A.B.B. and C.A.M. are responsible for the preparation of the paper. D.J.E., X.L., M.D.C., W.M., B.R. and K.A.B. are responsible for the experiments described in this paper.
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Eichelsdoerfer, D., Liao, X., Cabezas, M. et al. Large-area molecular patterning with polymer pen lithography. Nat Protoc 8, 2548–2560 (2013). https://doi.org/10.1038/nprot.2013.159
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DOI: https://doi.org/10.1038/nprot.2013.159
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