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用于高通量比色测定的醋酸纤维素微孔板。

Cellulose acetate microwell plates for high-throughput colorimetric assays.

作者信息

Gomez-Dopazo Gabriela B, Agosto Nieves Renis J, Albarracín Rivera Rolando L, Colon Morera Shaneily M, Nazario Daniel Rivera, Ramos Idalia, Dmochowski Ivan J, Lee Daeyeon, Bansal Vibha

机构信息

Department of Chemistry, University of Puerto Rico at Cayey 205 Ave. Antonio R Barcelo Cayey PR-00736 USA

Department of Physics and Electronics, University of Puerto Rico at Humacao CUH Station, 100 Road 908 Humacao PR-00791 USA.

出版信息

RSC Adv. 2024 May 13;14(22):15319-15327. doi: 10.1039/d4ra01317d. eCollection 2024 May 10.

Abstract

Single use plasticware (SUP) in scientific, diagnostic, and academic laboratories makes a significant contribution to plastic waste generation worldwide. Polystyrene (PS) microwell plates form a part of this waste. These plates are the backbone of high throughput colorimetric measurements in academic, research, and healthcare settings for detection/quantification of wide-ranging analytes including proteins, carbohydrates, nucleic acids, and enzyme activity. Polystyrene (PS) microwell plates serve as a platform for holding samples and reagents, where mixing initiates chemical reaction(s), and the ensuing color changes are quantified using a microplate reader. However, these plates are rarely reused or recycled, contributing to the staggering amounts of plastic waste generated in scientific laboratories. Here, we are reporting the fabrication of cellulose acetate (CA) microwell plates as a greener alternative to non-biodegradable PS plates and we demonstrate their application in colorimetric assays. These easy to fabricate, lighter weight, customizable, and environmentally friendly plates were fabricated in 96- and 384-well formats and made water impermeable through chemical treatment. The plates were tested in three different colorimetric analyses: (i) bicinchoninic acid assay (BCA) for protein quantification; (ii) chymotrypsin (CT) activity assay; and (iii) alkaline phosphatase (AP) activity assay. Color intensities were quantified using a freely available smartphone application, Spotxel® Reader (Sicasys Software GmbH). To benchmark the performance of this platform, the same assays were performed in commercial PS plates too and quantified using a UV/Vis microplate reader. The two systems yielded comparable linear correlation coefficients, LOD and LOQ values, thereby validating the CA plate-cell phone based analytical method. The CA microwell plates, coupled with smart phone optical data capture, provide greener, accessible, and scalable tools for all laboratory settings and are particularly well-suited for resource- and infrastructure-limited environments.

摘要

科学、诊断和学术实验室中的一次性塑料制品(SUP)对全球塑料垃圾的产生有重大影响。聚苯乙烯(PS)微孔板是此类垃圾的一部分。这些微孔板是学术、研究和医疗环境中高通量比色测量的核心,用于检测/定量包括蛋白质、碳水化合物、核酸和酶活性在内的各种分析物。聚苯乙烯(PS)微孔板用作容纳样品和试剂的平台,在该平台上混合引发化学反应,随后使用微孔板读数器对产生的颜色变化进行定量。然而,这些微孔板很少被重复使用或回收,导致科学实验室产生大量塑料垃圾。在此,我们报告了醋酸纤维素(CA)微孔板的制备,作为不可生物降解的PS板的更环保替代品,并展示了它们在比色分析中的应用。这些易于制造、重量更轻、可定制且环保的微孔板采用96孔和384孔格式制造,并通过化学处理使其不透水。这些微孔板在三种不同的比色分析中进行了测试:(i)用于蛋白质定量的二辛可宁酸(BCA)测定;(ii)胰凝乳蛋白酶(CT)活性测定;(iii)碱性磷酸酶(AP)活性测定。使用免费的智能手机应用程序Spotxel® Reader(Sicasys Software GmbH)对颜色强度进行定量。为了衡量该平台的性能,同样的分析也在商业PS板上进行,并使用紫外/可见微孔板读数器进行定量。这两个系统产生了可比的线性相关系数、检测限和定量限数值,从而验证了基于CA板 - 手机的分析方法。CA微孔板与智能手机光学数据采集相结合,为所有实验室环境提供了更环保、可及且可扩展的工具,尤其适用于资源和基础设施有限的环境。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac2f/11089462/dc17ed10f122/d4ra01317d-f1.jpg

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