Compact wide-field femtoliter-chamber imaging system for high-speed and accurate digital bioanalysis.
Lab Chip
; 23(4): 684-691, 2023 02 14.
Article
in En
| MEDLINE
| ID: mdl-36255223
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 â¼ 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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
COVID-19 Testing
/
COVID-19
Limits:
Humans
Language:
En
Journal:
Lab Chip
Journal subject:
BIOTECNOLOGIA
/
QUIMICA
Year:
2023
Document type:
Article
Affiliation country:
Japan
Country of publication:
United kingdom