Selective detection and automated counting of fluorescently-labeled chrysotile asbestos using a dual-mode high-throughput microscopy (DM-HTM) method

使用双模式高通量显微镜 (DM-HTM) 方法对荧光标记的温石棉进行选择性检测和自动计数

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作者:Myoung-Ock Cho, Hyo Mi Chang, Donghee Lee, Yeon Gyu Yu, Hwataik Han, Jung Kyung Kim

Abstract

Phase contrast microscopy (PCM) is a widely used analytical method for airborne asbestos, but it is unable to distinguish asbestos from non-asbestos fibers and requires time-consuming and laborious manual counting of fibers. Previously, we developed a high-throughput microscopy (HTM) method that could greatly reduce human intervention and analysis time through automated image acquisition and counting of fibers. In this study, we designed a dual-mode HTM (DM-HTM) device for the combined reflection and fluorescence imaging of asbestos, and automated a series of built-in image processing commands of ImageJ software to test its capabilities. We used DksA, a chrysotile-adhesive protein, for selective detection of chrysotile fibers in the mixed dust-free suspension of crysotile and amosite prepared in the laboratory. We demonstrate that fluorescently-stained chrysotile and total fibers can be identified and enumerated automatically in a high-throughput manner by the DM-HTM system. Combined with more advanced software that can correctly identify overlapping and branching fibers and distinguish between fibers and elongated dust particles, the DM-HTM method should enable fully automated counting of airborne asbestos.

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