Chemical imaging of biological systems with the scanning electrochemical microscope

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Abstract

A brief overview on recent advances in the application of scanning electrochemical microscopy (SECM) to the investigation of biological systems is presented. Special emphasis is given to the mapping of local enzyme activity by SECM, which is exemplified by relevant original systems.

Section snippets

Introduction, an overview

One of the most important and challenging area of analytical chemistry is the in-vivo or in-vitro collection of chemical information from targets of biological origin. The heterogeneity of biological systems requires in many cases a localized, small area measurement of specific analytes that eventually permits a better understanding of the function of the living organs investigated. The obvious spatial prerequisite is often coupled with high temporal resolution and minimal perturbation of the

Chemicals and reagents

Acetylcholinesterase enzyme (EC 3.1.1.7, from electric eel, type III as a solution containing 5 mg of ammonium sulfate per mg of protein), glucose oxidase (EC 1.1.3.4. from Aspergillus niger), acetylcholine chloride, glucose, bovine serum albumin, cystamine dihydrochloride and glutaraldehyde were purchased from Sigma (St. Louis, MS, USA). The 18-mer amino and thiol modified complementary oligonucleotids, with analogous sequence to the one located in the 662.-645. position of the Escherichia coli

Hybridization and enzyme labeling of the oligonucleotides

Four μl (1.8 nmol) of the complementary oligonucleotide strand solution (prepared with 0.1 M phosphate buffer, pH=7.4) was dropped onto the oligonucleotide modified surface and then 15 min was allowed for the hybridization. To detect the hybridization, the complementary oligonucleotide strand was covalently labeled with GOx. Therefore after hybridization, the surface of the UMEA was thoroughly rinsed and then contacted for 1 h with 2.5% glutaraldehyde solution to react with the amino group of

SECM imaging

SECM images were recorded with a laboratory made scanning electrochemical microscope based on three high resolution stepper motor driven translation stages (Newport, Evry, France) and a PGSTAT 10 (Ecochemie, Utrecht, The Netherlands) equipped with a EDC preamplifier for low current measurements. The data acquisition and the positioning system were controlled by self-developed software operating on a Pentium II personal computer. For potentiometric measurements a custom designed high input

Imaging of biological systems by scanning potentiometric microscopy

The use of ion-selective potentiometry in SECM offers a selective detection technique for ions that are difficult to assess by voltammetric means, such as calcium, hydrogen, sodium, ammonium, etc. In contrast with voltammetric tips, potentiometric microelectrodes are passive tips, which mean that the detection does not require the conversion of the detected component at the electrode and feedback effects cannot be generated. These particularities of the potentiometric operation mode can

Conclusion

The overview on the application of SECM to biological systems and the selected original examples clearly demonstrates the potential of scanning electrochemical microscopy in the field of biology and biomedical science. The technique proved to be lately very useful for controlling the preparation and the characterization of biosensors and sensor structures, as well as the optimization of their performance.

Acknowledgements

This work was financially supported by the Hungarian National Science Foundation (OTKA M027974, OTKA M041969, OTKA F037977) and the strategic grant of the Budapest University of Technology and Economics (Nr: 26276). R. E. Gyurcsányi gratefully acknowledges the Bolyai János and Varga József (Balla György) fellowships. We thank Dr. E. Lindner for generously providing the microelectrode arrays indispensable for the enzyme patterning experiments and for inspiring discussions.

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