FluoroPi Device With SmartProbes: A Frugal Point-of-Care System for Fluorescent Detection of Bacteria From a Pre-Clinical Model of Microbial Keratitis.
FluoroPi Device With SmartProbes: A Frugal Point-of-Care System for Fluorescent Detection of Bacteria From a Pre-Clinical Model of Microbial Keratitis.
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DOI：
10.1167/tvst.12.7.1
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发表时间：
2023-07-03
影响因子：
3
通讯作者：
Mills, Bethany
中科院分区：
文献类型：
--
作者：
Mohanan, Syam Mohan P. C.;Russell, Kay;Duncan, Sheelagh;Kiang, Alex;Lochenie, Charles;Duffy, Emma;Kennedy, Stephnie;Prajna, N. Venkatesh;Williams, Rachel L.;Dhaliwal, Kevin;Williams, Gareth O. S.;Mills, Bethany
快速准确地诊断微生物性角膜炎（MK）可以大大改善患者的预后。在这里，我们提出了一种快速，可访问的多色荧光成像设备（FluoroPi）的开发，并结合荧光光学报告（SmartProbes）来区分细菌革兰氏状态，以评估其性能。此外，我们通过对来自离体猪角膜MK模型的角膜刮擦和微创角膜印模膜（CIM）获得的样本进行成像显示了可行性。FluoroPi使用Raspberry Pi单板计算机和相机、发光二极管（LED）以及用于白光和荧光成像的滤光片构建，并激发和检测细菌光学SmartProbes：革兰氏阴性，NBD-PMX（exmax 488 nm）;革兰氏阳性，Merocy-Van（exmax 590 nm）。我们用细菌（铜绿假单胞菌和金黄色葡萄球菌）评价了FluoroPi，这些细菌是通过刮擦（针）从MK的离体猪角膜模型中分离的，并用SmartProbes评价了CIM。FluoroPi提供<1 µm的分辨率，并且当与SmartProbes结合时，能够容易地区分从MK的离体模型分离的细菌和组织碎片，通过刮擦和CIM检索。单个细菌可在视野内分辨，检测限为103至104 CFU/mL。成像前的样品制备非常简单（免清洗），使用FluoroPi进行成像和后处理非常简单，证实了其易用性。FluoroPi与SmartProbes结合可提供有效、低成本的细菌成像，描绘直接从MK临床前模型中采样的革兰氏阴性和革兰氏阳性细菌。这项研究为MK的快速，微创诊断方法的临床转化提供了关键的垫脚石。
Rapid and accurate diagnosis of microbial keratitis (MK) could greatly improve patient outcomes. Here, we present the development of a rapid, accessible multicolour fluorescence imaging device (FluoroPi) and evaluate its performance in combination with fluorescent optical reporters (SmartProbes) to distinguish bacterial Gram status. Furthermore, we show feasibility by imaging samples obtained by corneal scrape and minimally invasive corneal impression membrane (CIM) from ex vivo porcine corneal MK models. FluoroPi was built using a Raspberry Pi single-board computer and camera, light-emitting-diodes (LEDs), and filters for white-light and fluorescent imaging, with excitation and detection of bacterial optical SmartProbes: Gram-negative, NBD-PMX (exmax 488 nm); Gram positive, Merocy-Van (exmax 590 nm). We evaluated FluoroPi with bacteria (Pseudomonas aeruginosa and Staphylococcus aureus) isolated from ex vivo porcine corneal models of MK by scrape (needle) and CIM with the SmartProbes. FluoroPi provides <1 µm resolution and was able to readily distinguish bacteria isolated from ex vivo models of MK from tissue debris when combined with SmartProbes, retrieved by both scrape and CIM. Single bacteria could be resolved within the field of view, with limits of detection demonstrated as 103 to 104 CFU/mL. Sample preparation prior to imaging was minimal (wash-free), and imaging and postprocessing with FluoroPi were straightforward, confirming ease of use. FluoroPi coupled with SmartProbes provides effective, low-cost bacterial imaging, delineating Gram-negative and Gram-positive bacteria directly sampled from a preclinical model of MK. This study provides a crucial stepping stone toward clinical translation of a rapid, minimally invasive diagnostic approach for MK.
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DOI：
10.1007/s10856-019-6303-1
发表时间：
2019-09-01
影响因子：
3.7
作者：
Kennedy, Stephnie;Lace, Rebecca;Levis, Hannah J.
通讯作者：
Levis, Hannah J.
影响因子：
4.6
作者：
Orth A;Wilson ER;Thompson JG;Gibson BC
通讯作者：
Gibson BC
影响因子：
2.7
作者：
Low L;Fuentes-Utrilla P;Hodson J;O'Neil JD;Rossiter AE;Begum G;Suleiman K;Murray PI;Wallace GR;Loman NJ;Rauz S;West Midlands Collaborative Ophthalmology Network for Clinical Effectiveness & Research by Trainees (WM CONCERT)
通讯作者：
West Midlands Collaborative Ophthalmology Network for Clinical Effectiveness & Research by Trainees (WM CONCERT)
影响因子：
1.8
作者：
Chidambaram, Jaya Devi;Prajna, Namperumalsamy Venkatesh;Burton, Matthew J.
通讯作者：
Burton, Matthew J.
影响因子：
3.7
作者：
Granot Y;Ivorra A;Rubinsky B
通讯作者：
Rubinsky B