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通过硝酸纤维素膜中的激光微加工约束实现侧向流动分析的灵敏度和信号增强。

Lateral flow assay sensitivity and signal enhancement via laser µ-machined constrains in nitrocellulose membrane.

作者信息

Khatmi Gazy, Klinavičius Tomas, Simanavičius Martynas, Silimavičius Laimis, Tamulevičienė Asta, Rimkutė Agnė, Kučinskaitė-Kodzė Indrė, Gylys Gintautas, Tamulevičius Tomas

机构信息

Department of Physics, Kaunas University of Technology, Kaunas, Lithuania.

Institute of Materials Science, Kaunas University of Technology, Kaunas, Lithuania.

出版信息

Sci Rep. 2024 Oct 2;14(1):22936. doi: 10.1038/s41598-024-74407-3.

Abstract

Lateral flow assay (LFA) is a handful diagnostic technology that can identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other common respiratory viruses in one strip, which can be tested at the point-of-care without the need for equipment or skilled personnel outside the laboratory. Although its simplicity and practicality make it an appealing solution, it remains a grand challenge to substantially enhance the colorimetric LFA sensitivity. In this work, we present a straightforward approach to enhance the sensitivity of LFA by imposing the flow constraints in nitrocellulose (NC) membranes via a number of vertical femtosecond laser micromachined microchannels which is important for prolonged specific binding interactions. Porous NC membrane surfaces were structured with different widths and densities µ-channels employing a second harmonic of the Yb:KGW femtosecond laser and sample XYZ translation over a microscope objective-focused laser beam. The influence of the microchannel parameters on the vertical wicking speed was evaluated from the video recordings. The obtained results indicated that µ-channel length, width, and density in NC membranes controllably increased the immunological reaction time between the analyte and the labeled antibody by 950%. Image analysis of the colorimetric indicators confirmed that the flow rate delaying strategy enhanced the signal sensitives by 40% compared with pristine NC LFA.

摘要

侧向流动分析(LFA)是一种便捷的诊断技术,能够在一条试纸条上识别严重急性呼吸综合征冠状病毒2(SARS-CoV-2)和其他常见呼吸道病毒,无需实验室外的设备或专业人员即可在护理点进行检测。尽管其简单实用性使其成为一个有吸引力的解决方案,但大幅提高比色LFA的灵敏度仍然是一个巨大的挑战。在这项工作中,我们提出了一种直接的方法来提高LFA的灵敏度,即通过许多垂直的飞秒激光微加工微通道在硝酸纤维素(NC)膜中施加流动限制,这对于延长特异性结合相互作用很重要。采用Yb:KGW飞秒激光的二次谐波以及样品在显微镜物镜聚焦激光束上的XYZ平移,在多孔NC膜表面构建了不同宽度和密度的微通道。从视频记录中评估了微通道参数对垂直毛细上升速度的影响。所得结果表明,NC膜中的微通道长度、宽度和密度可将分析物与标记抗体之间的免疫反应时间可控地延长950%。比色指示剂的图像分析证实,与原始NC LFA相比,流速延迟策略使信号灵敏度提高了40%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab73/11446913/94b832bb2f4f/41598_2024_74407_Fig1_HTML.jpg

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