Abstract
Oxygen plasma micro-nanostructured poly(methyl methacrylate) (PMMA) slides were modified through silver microparticle deposition to create microarray substrates that enhance the emitted fluorescence intensity. Silver deposition relied on a commercially available reagent and was completed in two 30-min incubation cycles of the substrate with the reagent. The fluorescence enhancement achieved using these substrates over flat PMMA slides was determined through the development of a microarray for the multiplexed detection of four mycotoxins, aflatoxin B1, ochratoxin A, fumonisin B1, and deoxynivalenol. It was shown that the implementation of silver-plated oxygen plasma micro-nanotextured PMMA substrates increased the signals obtained for aflatoxin B1 and ochratoxin A by approximately 2.8 times, 5.6 times for deoxynivalenol, and 16-times for fumonisin B1, compared to flat PMMA substrates. Most notably, this signal increase was not accompanied by a significant increase in the non-specific signal. In addition, the spot repeatability both across a single slide as well as between different slides was high, with coefficients of variation lower than 12%. The slides were also stable for at least three months, thus offering a microarray substrate with improved properties compared to standard glass slides, regarding both the absolute spot fluorescence intensity and between spots repeatability.
Highlights
Mycotoxins are low molecular weight compounds, produced as secondary metabolites by fungi, that contaminate agricultural products during cultivation, harvest and post-harvest procedures and can have toxic effects on the livestock and humans that consume the contaminated commodities [1]
3D micro-nanostructured polymer films spin-coated over conventional glass slides, which have been previously employed as microarray substrates [16,17] were implemented for preparation of metal-coated 3D substrates using a commercially available silver enhancement kit
Nucleation is very limited in absence of gold nanoparticles, from preliminary experiments it was found that nucleation and Ag deposition is possible on both flat, as well as on micro-nanotextured surfaces, especially when repetitive cycles of silver enhancement solution application followed by washings were applied
Summary
Mycotoxins are low molecular weight compounds, produced as secondary metabolites by fungi, that contaminate agricultural products during cultivation, harvest and post-harvest procedures and can have toxic effects on the livestock and humans that consume the contaminated commodities [1]. Mycotoxins are known for their acute and chronic toxic, immunosuppressive, mutagenic, teratogenic, or even carcinogenic effects [3]. These effects have generally been studied for individual mycotoxins or classes thereof, despite the fact that several mycotoxins belonging to different classes might be found in a single product [4,5]. Amongst the various mycotoxins of concern, with regard to effects on human health, are aflatoxins, ochratoxins, fumonisins, and trichothecenes [1], and the main representatives of these classes are aflatoxin B1 (AFB1), ochratoxin A (OTA), fumonisin B1 cence approach with the plasma 3D micro-nanostructured surfaces in order microarray substrates that can combine the virtues of both technologies and means for the development of highly sensitive fluorescence microarrays. The substrates (FB1), order taonpdpcrodhltoeeeoymcxetyidbcnaoifltvohdarlteehttnheeocehltie(foDaanlbOthroNifco)aff,ottriuheoesrnpmpeoucyftbiclvmoicetliobycxur(oitFnaaisgrl,rsunaoryaetsmh1e)fe.oflayrItrmAtihsFe,erBtshs1eai,mrnOedfuTolfrAtoeao,,nddFeeBops1urior,sadabumnlcedueirlDnstiOplNex, eind by unnecepsslaersyspinrocedutcheesreecafollus,rtomdyectoetcotxminusltiaprlee mmoycsot tforxeiqnus eonftdlyiffeenrecnotucnlatessreesdininacereal g single run.feeds (corn, oat, wheat, barley, etc.) [26]
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