Hydrogel-enzyme micropatch array format for chemical mapping: A proof of concept.

Autor: Huang LL; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan., Chua ZQ; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan., Buchowiecki K; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan., Raju CM; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan., Urban PL; Department of Chemistry, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan; Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University, 101, Section 2, Kuang-Fu Rd., Hsinchu, 300044, Taiwan. Electronic address: urban@mx.nthu.edu.tw.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2023 Nov 01; Vol. 239, pp. 115599. Date of Electronic Publication: 2023 Aug 14.
DOI: 10.1016/j.bios.2023.115599
Abstrakt: Conventional sensing methods report on concentrations of analytes in a single point of sampled medium or provide an average value. However, distributions of substances on surfaces of sampled objects often exhibit intricate inhomogeneities. In order to obtain snapshots of the chemical distributions on surfaces, we have developed enzyme-loaded hydrogel arrays (5 × 5 and 10 × 10). The acrylic 10 × 10 array base contains 100 holes, which are filled with agarose hydrogel containing assay enzymes and substrates. Such arrays can be exposed to the analyzed surfaces to collect minute amounts of analytes. Following a brief incubation, they are subsequently visualized in a custom-built array reader device. The reader incorporates a light-emitting diode-based light source, miniature camera, and Raspberry Pi single-board computer. Two Python programs capture and analyze the images of the array to extract pixel saturation values corresponding to individual hydrogel micropatches. The method has been thoroughly optimized for mapping of glucose and lactic acid. The optimized parameters were: contact time, agarose concentration, substrate concentration, enzyme concentration ratio, and enzyme concentration. The array biosensor was further tested by mapping glucose distribution in fruit/vegetable cross-sections (apple, guava, and cucumber) and lactic acid distribution in cheese. We think that this new hydrogel-based chemical mapping method can find applications in studies related to food science, plant physiology, clinical chemistry, and forensics; wherever the distributions of analytes on the tested surfaces need to be assessed.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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