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Real-time detection of an airborne microorganism using inertial impaction and mini-fluorescent microscopy

机译:使用惯性碰撞和微型荧光显微镜实时检测空气传播的微生物

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摘要

To achieve successful real-time detection of airborne pathogenic microorganisms, the problem must be considered in terms of their physical size and biological characteristics. We developed an airborne microorganism detection chip to realize the detection of microorganisms, ensuring compactness, sensitivity, cost-efficiency, and portability, using three key components: an inertial impaction system, a cartridge-type impaction plate, and a mini-fluorescent microscope. The inertial impaction system was used to separate microorganisms in terms of their aerodynamic particle size, and was fabricated with three impaction stages. Numerical analysis was performed to design the system; the calculated cutoff diameter at each impaction stage was 2.02 (first stage), 0.88 (second stage), and 0.54 μm (third stage). The measured cutoff diameters were 2.24, 0.91, and 0.49 μm, respectively. A cartridge-type impaction plate was used, composed of molded polydimethylsiloxane (PDMS) and an actual impaction region made of a SYBR green I dye-stained agar plate. A mini-fluorescent microscope was used to distinguish microbes from non-biological particles. Images of the microorganisms deposited at the impaction zone were obtained via mini-fluorescent microscopy, and fluorescent intensities of the images were calculated using in-house image-processing software. The results showed that the developed system successfully identified aerosolized biological particles from non-biological particles in real time.
机译:为了成功地实时检测出空气传播的病原微生物,必须根据其物理大小和生物学特性来考虑该问题。我们开发了一种机载微生物检测芯片,它使用三个关键组件实现了对微生物的检测,从而确保了紧凑性,灵敏性,成本效益和便携性:惯性撞击系统,盒式撞击板和微型荧光显微镜。惯性碰撞系统用于根据微生物的空气动力学粒径对其进行分离,并由三个碰撞阶段组成。进行了数值分析以设计系统;在每个撞击阶段,计算出的截止直径分别为2.02(第一阶段),0.88(第二阶段)和0.54μm(第三阶段)。测得的截止直径分别为2.24、0.91和0.49μm。使用的盒式冲击板由模制的聚二甲基硅氧烷(PDMS)组成,实际的冲击区域由SYBR green I染色的琼脂板制成。微型荧光显微镜用于区分微生物和非生物颗粒。通过微型荧光显微镜获得沉积在撞击区域的微生物的图像,并使用内部图像处理软件计算图像的荧光强度。结果表明,开发的系统成功地从非生物颗粒中实时识别了气溶胶化的生物颗粒。

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