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Integration of an electrochemical-based biolithography technique into an AFM system

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Abstract

An ordinary atomic force microscopy (AFM) was functionalized and applied to electrochemically draw micropatterns of biomolecules. To fabricate an electrochemical AFM probe having an electrode at the tip, a metal-coated AFM probe was first insulated with Parylene C, and then the apex of the tip was ground mechanically to expose the electrode. The effective electrode diameter was estimated to be ca. 500 nm. The electrode probe was positioned close to a heparin-coated antibiofouling substrate and used to locally generate hypobromous acid from a dilute Br− solution to render the substrate surface protein-adhesive. In situ topographical imaging after the electrochemical treatment suggested the heparin layer became detached to allow the adsorption of proteins, in this case fibronectin. The diameter of the drawn fibronectin pattern was 2 μm, which is one order of magnitude smaller than we achieved previously using a microdisk electrode (tip diameter 10 μm).

AFM configuration integrated with the electrochemical-based surface modification and resultant micropatterns of fluorescence-labeled fibronectin

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Acknowledgements

This study was partially supported by the Industrial Technology Research Grant Program from NEDO of Japan, by Grants-in-Aid for Scientific Research B (No. 17310080), for Scientific Research on Priority Areas (No. 18048004), and for Young Scientists (startup) (No. 18810004) from the Ministry of Education, Science, and Culture, Japan, and by grants from the “Hattori-Hokokai” Foundation and the Advanced Technology Institute Foundation.

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Authors and Affiliations

  1. Department of Bioengineering and Robotics, Graduate School of Engineering, Tohoku University, Sendai, 980–8579, Japan

    Soichiro Sekine, Hirokazu Kaji & Matsuhiko Nishizawa

Authors
  1. Soichiro Sekine
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  2. Hirokazu Kaji
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  3. Matsuhiko Nishizawa
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Corresponding authors

Correspondence to Hirokazu Kaji or Matsuhiko Nishizawa.

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Suppl. Fig. 1

Cyclic voltammogram of a Pt disk electrode (tip diameter 10 μm) measured in the two-electrode configuration with an Ag/AgCl counter electrode in PBS containing 25 mM KBr at a scan rate of 100 mV s−1. Since the electrochemical oxidation of Br− is controlled by charge-transfer processes, the oxidation current is proportional to the surface area of the electrode. The observed oxidation current at 1.7 V is ca. 300 nA, which is 500 times higher than the current value obtained by using the probe tip electrode (red line in Fig. 3b). Assuming that the probe tip electrode is disk-type, its diameter is estimated to be ca. 500 nm. (DOC 75.0 kb)

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Sekine, S., Kaji, H. & Nishizawa, M. Integration of an electrochemical-based biolithography technique into an AFM system. Anal Bioanal Chem 391, 2711–2716 (2008). https://doi.org/10.1007/s00216-008-1952-9

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  • DOI: https://doi.org/10.1007/s00216-008-1952-9

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