Journal of Applied Optics, Volume. 44, Issue 4, 859(2023)
Multiple real-time fluorescence detection system for rapid on-site nucleic acid detection
Figures & Tables(17)
Fig. 3. Comparison diagram of emission optical path simulation and illumination
Fig. 7. Test results of fluorescence dye gradient for each channel
Table 1. Semi-divergence angle measured at each end of optical path
（°） View tableView in ArticleTable 1. Semi-divergence angle measured at each end of optical path
（°） 位置 激发光路前端 芯片激发端 发射光路前端 发散角 19.03 13.50 18.78
Table 2. Distance between elements of emission optical path
mm View tableView in ArticleTable 2. Distance between elements of emission optical path
mm 距离 发射光纤-透镜 透镜-发射滤光片 发射滤光片-PD 优化前距离 11.8 3.7 3.1 优化后距离 8.8 5.8 2.5
Table 3. Dilution concentration of fluorescent dyes
μg·mL−1 View tableView in ArticleTable 3. Dilution concentration of fluorescent dyes
μg·mL−1 荧光染料 FAM HEX ROX CY5 母液（50%DMSO） 2 000 13 200 18 梯度1（DEPC水） 200 1.3 20 1.8 梯度2（DEPC水） 100 0.65 10 0.9 梯度3（DEPC水） 50 0.325 5 0.45 梯度4（DEPC水） 25 0.162 5 2.5 0.225 梯度5（DEPC水） 12.5 0.081 25 1.25 0.112 5
Table 4. Detection limit test results of fluorescent dyes
View tableView in ArticleTable 4. Detection limit test results of fluorescent dyes
检测通道 ROX FAM HEX CY5 检测下限/μg·mL−1 0.31 1.56 0.02 0.11 线性度R2值 0.9961 0.9472 0.9968 0.9935
Table 5. Gradient test results of fluorescent dyes
View tableView in ArticleTable 5. Gradient test results of fluorescent dyes
检测通道 电流值对数 线性度R2值 梯度5 梯度4 梯度3 梯度2 梯度1 ROX 2.99 3.88 4.84 5.88 6.84 0.999 3 FAM 1.70 2.21 3.22 4.17 5.01 0.990 7 HEX 5.67 6.54 7.43 8.61 9.39 0.996 5 CY5 4.61 5.27 6.10 6.87 7.39 0.994 8
Table 6. Repeatability test results of fluorescence intensity
View tableView in ArticleTable 6. Repeatability test results of fluorescence intensity
检测通道 CV值 梯度5/% 梯度3/% 梯度1/% ROX 1.27 0.19 0.16 FAM 1.20 0.89 0.59 HEX 1.01 0.27 0.06 CY5 1.24 0.20 0.29
Table 7. Fluorescence crosstalk test results based on HEX fluorescent dyes
pA View tableView in ArticleTable 7. Fluorescence crosstalk test results based on HEX fluorescent dyes
pA 检测通道 电流值 高浓度 中浓度 低浓度 DEPC水 ROX 1.90 1.11 2.15 1.28 FAM 0.29 0.54 1.23 0.22 HEX 217.71 65.63 20.52 4.18 CY5 0.80 0.42 0.67 1.19
Table 8. Fluorescence crosstalk test results based on CY5 fluorescent dyes
pA View tableView in ArticleTable 8. Fluorescence crosstalk test results based on CY5 fluorescent dyes
pA 检测通道 电流值 高浓度 中浓度 低浓度 DEPC水 ROX 0.22 0.37 0.67 1.15 FAM 0.28 0.14 0.19 0.02 HEX 2.03 0.19 1.78 1.37 CY5 120.63 38.54 14.15 0.94
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Jiayu YANG, Juntian ZENG, Bangchao XI, Guoqiang LIU, Shaolei HUANG, Dongxu ZHANG. Multiple real-time fluorescence detection system for rapid on-site nucleic acid detection[J]. Journal of Applied Optics, 2023, 44(4): 859
Paper Information
Category: Research Articles
Received: Sep. 29, 2022
Accepted: --
Published Online: Aug. 10, 2023
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