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Compact wide-field femtoliter-chamber imaging system for high-speed and accurate digital bioanalysis
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-10-18 , DOI: 10.1039/d2lc00741j
Tatsuya Iida 1 , Jun Ando 1 , Hajime Shinoda 1 , Asami Makino 1 , Mami Yoshimura 1 , Kazue Murai 1 , Makiko Mori 1 , Hiroaki Takeuchi 2 , Takeshi Noda 3 , Hiroshi Nishimasu 4, 5 , Rikiya Watanabe 1
Affiliation  

The femtoliter-chamber array is a bioanalytical platform that enables highly sensitive and quantitative analysis of biological reactions at the single-molecule level. This feature has been considered a key technology for “digital bioanalysis” in the biomedical field; however, its versatility is limited by the need for a large and expensive setup such as a fluorescence microscope, which requires a long time to acquire the entire image of a femtoliter-chamber array. To address these issues, we developed a compact and inexpensive wide-field imaging system (COWFISH) that can acquire fluorescence images with a large field of view (11.8 mm × 7.9 mm) and a high spatial resolution of ∼ 3 μm, enabling high-speed analysis of sub-million femtoliter chambers in 20 s. Using COWFISH, we demonstrated a CRISPR-Cas13a-based digital detection of viral RNA of SARS-CoV-2 with an equivalent detection sensitivity (limit of detection: 480 aM) and a 10-fold reduction in total imaging time, as compared to confocal fluorescence microscopy. In addition, we demonstrated the feasibility of COWFISH to discriminate between SARS-CoV-2-positive and -negative clinical specimens with 95% accuracy, showing its application in COVID-19 diagnosis. Therefore, COWFISH can serve as a compact and inexpensive imaging system for high-speed and accurate digital bioanalysis, paving a way for various biomedical applications, such as diagnosis of viral infections.

中文翻译:

用于高速准确数字生物分析的紧凑型宽视场飞升腔室成像系统

femtoliter-chamber array 是一个生物分析平台,可以在单分子水平上对生物反应进行高灵敏度和定量分析。这一特性被认为是生物医学领域“数字化生物分析”的关键技术;然而,它的多功能性受到需要大型且昂贵的设置(例如荧光显微镜)的限制,这需要很长时间才能获取飞升腔室阵列的整个图像。为了解决这些问题,我们开发了一种紧凑且廉价的宽视场成像系统 (COWFISH),它可以获取具有大视场 (11.8 mm × 7.9 mm) 和 ∼ 3 μm 高空间分辨率的荧光图像,从而实现高-在 20 秒内对亚百万飞升腔室进行速度分析。使用 COWFISH,与共聚焦荧光显微镜相比,我们展示了一种基于 CRISPR-Cas13a 的 SARS-CoV-2 病毒 RNA 数字检测,具有等效的检测灵敏度(检测限:480 aM)并且总成像时间减少了 10 倍。此外,我们证明了 COWFISH 以 95% 的准确率区分 SARS-CoV-2 阳性和阴性临床标本的可行性,展示了其在 COVID-19 诊断中的应用。因此,COWFISH 可以作为一种紧凑、廉价的成像系统,用于高速、准确的数字生物分析,为各种生物医学应用铺平道路,例如病毒感染的诊断。我们证明了 COWFISH 以 95% 的准确率区分 SARS-CoV-2 阳性和阴性临床标本的可行性,展示了其在 COVID-19 诊断中的应用。因此,COWFISH 可以作为一种紧凑、廉价的成像系统,用于高速、准确的数字生物分析,为各种生物医学应用铺平道路,例如病毒感染的诊断。我们证明了 COWFISH 以 95% 的准确率区分 SARS-CoV-2 阳性和阴性临床标本的可行性,展示了其在 COVID-19 诊断中的应用。因此,COWFISH 可以作为一种紧凑、廉价的成像系统,用于高速、准确的数字生物分析,为各种生物医学应用铺平道路,例如病毒感染的诊断。
更新日期:2022-10-18
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