22 February 2013 A preliminary investigation on the interaction between sol-gel immobilized glucose oxidase and freely diffusing glucose by means of two-photon microscopy
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Abstract
To study immobilized protein interactions with dissolved substrates is a very important topic both from a fundamental and technological standpoint. In the present report we illustrate the preliminary results obtained on sol-gel immobilized glucose oxidase (GOD) using a standard de-scanned two-photon microscope based on a modified confocal scanhead with internal detectors and a Ti:sapphire laser as a source. Data acquisition conditions were preliminary defined using functionalized beads of different dimensions. Various sol-gel supports were then investigated by monitoring endogeneous fluorescence due to the flavoadenine (FAD) molecules, present in GOD. Linear absorption and fluorescence spectroscopy along with Fourier Transform Infrared microscopy were employed for a full-optical characterization of the samples. The results show that GOD immobilization processes can be successfully monitored in some cases and also the interaction with glucose could be studied by this approach. This assessment holds potentials to better understand the characteristic of immobilized enzymes biocatalysis and to develop new biosensing schemes.
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I. Delfino, I. Delfino, M. Portaccio, M. Portaccio, M. De Rosa, M. De Rosa, M. Lepore, M. Lepore, } "A preliminary investigation on the interaction between sol-gel immobilized glucose oxidase and freely diffusing glucose by means of two-photon microscopy", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85882U (22 February 2013); doi: 10.1117/12.2006347; https://doi.org/10.1117/12.2006347
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