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제124회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Digital and Absolute Quantification of Microdroplets using Wide-Field Imaging System for real-time Droplet Sorting

2019년 8월 22일 11시 35분 09초
ANAL1.O-7 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
목 09시 : 18분
Analytical Chemistry - Oral Presentation of Young Analytical Chemists I
저자 및
Sunghyun Ki, Dong-Ku Kang1,*
Chemistry, Incheon National University, Korea
1Department of Chemistry, Incheon National University, Korea
Droplet-based microfluidics (DMF) have been developed as a digital assay that allow precise and absolute quantification of targets such as enzyme, bacteria and nucleic acid. To avoid complex and expensive optic system as a detector, CCD sensors and smartphones have been integrated with DMF to image droplets. Here, we introduce digital CFU assay for absolute quantification of living-bacteria using 50.6-megapixel CMOS sensor which allow imaging the area of 10 cm2. Detectable number of droplets depends on the diameter of droplets, and it can be calculated as 9.29×106, 5.8×105 or 1.5×105 droplets when droplets are generated as 10, 40 or 80 μm in diameter, respectively. In this approach, 107 droplets can be analyzed within one minute after being generated and digital CFU assay was performed as a proof of concept study for the absolute quantification of living bacteria without fluorescent labeling. As a wide-field imaging system, 100 mm macro and full-frame 50.6-megapixel CMOS sensor-integrated digital camera were used to image droplets, and droplet images were analyzed using Matlab that simultaneously analyzes five individual colors. Bacterial cells (E.coli) were encapsulated within droplets as a single cell manner using microfluidic device and phenol-red was co-encapsulated as a pH indicator to detect living bacteria. In addition, we have investigated the migration of ammonia gas from bacteria and colorimetric dye between droplets. Interestingly, the dye was not able to across between the droplets but ammonia gas permeated through the droplets. Here, we introduce a detection platform (wide-field imaging system) that can be used for the monitoring large number of droplets (up to 107) within 5 minutes.