Spectroscopy and microscopy characterization of spiropyran autoassemble device

There are many studies focuses on self-assembled films characterization, photochromic material (spiropyran) had been electrosattached to different enzymes, and in this work we used these spiropyran characteristics to immobilized glucose oxidase. The aim of this work is establish a protocol to build an optical device that shows absorption changes when an enzyme is immobilized and characterized the devise through FTIR and UV/Vis spectroscopy and Atomic Force and Scanning Electron Microscopy. We use different methods such as self-assembled monolayers and the activation reaction by carbodiimide. Corning of quartz was used as matrix, clean solution (1:1 MeOH:HCl, H ₂SO₄) was used to prepared the matrix, 1% silane solution was prepared to functionalized the surface, and a solution prepared with spiropyran (C₁₉H₁₈N₂O₃), EDC (C5H11N=C=NC6H11), NHS (C₄H₅NO₃), HEPES (C ₈H₁₈N₂O₄S) and glucose oxidase from Aspergillus niger was the last step to immobilized this enzyme. Three steps to build it, the first step was cleaned the matrix, the next step was functionalized the surface with silane material, this step allows to modified the surfaces, to prepared to attach the spiropyran. The bonds found from the FTIR spectra to silanized step and the functionalized step (spiropyran attached). That indicated that FTIR and AFM techniques are available to characterized and identify absorption, morphology of a devise modified with spiropyran, that allows attach some biological material, in this case glucose oxidase, the absorption changes and morphological changes are the evidence of immobilization successful.