Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy
Abstract
:1. Introduction
2. Methods
2.1. Overall Micro-Droplet Detection
2.1.1. Noise Reduction with the Gaussian Filter
2.1.2. Contrast Limited Adaptive Histogram Equalization
2.1.3. Maximizing inter-class Variance Thresholding Method
2.1.4. Circle Detection via Circular Hough Transform
- Accumulator array computation:The edge detection is carried out on the binary map to get an edge image (L). The edge pixels of L are designated as candidate pixels and are allowed to cast ‘votes’ in the accumulator array A(a), which represents the weight of the circle with a fixed radius and the center of the circle. Here, . (a, b) represents the space location of pixels, and r is the radius of the expected circle. At the beginning, all the elements of A(a) are set to 0.
- Center and radius estimation:For every pixel x of the fluorescence image, we accumulate all the units of A(a) that satisfy the function . is the analytical expression of circle:Finally, the circular centers and radii are estimated by detecting the peaks in the accumulator array. We can get the number of micro-droplets by counting the centers of detected circles.
2.2. Fluorescent Micro-Droplet Detection
2.3. Measurement of AP-Labeled Nanoparticle Concentration
2.4. Evaluation
2.5. Code
3. Results
3.1. Overall Micro-Droplet Detection
3.1.1. Visual Evaluation
3.1.2. TPR and FPR
3.1.3. ROC and F-Measure
3.1.4. Detected Number of Overall Micro-Droplets
3.2. Fluorescent Micro-Droplet Detection
3.3. AP-Labeled Nanoparticle Concentration Measurement
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Samples | MSVST | MSSEF | MPHD | The Proposed Method |
---|---|---|---|---|
Image1 | 0.9204 | 0.7414 | 0.6640 | 0.9231 |
Image 2 | 0.9306 | 0.7889 | 0.7250 | 0.9674 |
Image 3 | 0.9348 | 0.8127 | 0.6610 | 0.9597 |
Image 4 | 0.9260 | 0.7678 | 0.6591 | 0.9656 |
Image 5 | 0.9075 | 0.8038 | 0.6392 | 0.9770 |
Image 6 | 0.9343 | 0.7737 | 0.7318 | 0.9677 |
Image 7 | 0.8945 | 0.7607 | 0.6311 | 0.9604 |
Image 8 | 0.8931 | 0.7564 | 0.6183 | 0.9663 |
Image 9 | 0.8792 | 0.7569 | 0.5946 | 0.9721 |
Image 10 | 0.8810 | 0.5775 | 0.6183 | 0.9402 |
Image 11 | 0.8999 | 0.6082 | 0.6653 | 0.9655 |
Image 12 | 0.8462 | 0.6044 | 0.5859 | 0.9186 |
Image 13 | 0.9177 | 0.6545 | 0.6586 | 0.9707 |
Image 14 | 0.9202 | 0.5954 | 0.6555 | 0.9692 |
Image 15 | 0.8831 | 0.6368 | 0.5987 | 0.9551 |
Average | 0.9046 | 0.7093 | 0.6471 | 0.9586 |
Samples | True Number | MSVST | MSSEF | MPHD | The Proposed Method |
---|---|---|---|---|---|
Image1 | 161 | 163 | 93 | 152 | 161 |
Image 2 | 222 | 232 | 142 | 202 | 222 |
Image 3 | 221 | 223 | 142 | 202 | 221 |
Image 4 | 223 | 227 | 135 | 198 | 222 |
Image 5 | 219 | 224 | 149 | 202 | 218 |
Image 6 | 229 | 235 | 152 | 210 | 229 |
Image 7 | 250 | 255 | 150 | 236 | 249 |
Image 8 | 239 | 245 | 149 | 224 | 240 |
Image 9 | 245 | 246 | 141 | 224 | 245 |
Image 10 | 381 | 393 | 155 | 350 | 381 |
Image 11 | 372 | 383 | 159 | 348 | 372 |
Image 12 | 381 | 386 | 175 | 345 | 381 |
Image 13 | 347 | 356 | 166 | 320 | 349 |
Image 14 | 414 | 422 | 175 | 371 | 412 |
Image 15 | 358 | 365 | 164 | 325 | 357 |
Samples | True Number | Detected Number of Fluorescent Micro-Droplets | Relative Error |
---|---|---|---|
Image1 | 21 | 21 | 0.00% |
Image 2 | 18 | 18 | 0.00% |
Image 3 | 18 | 18 | 0.00% |
Image 4 | 16 | 17 | 6.25% |
Image 5 | 13 | 13 | 0.00% |
Image 6 | 24 | 24 | 0.00% |
Image 7 | 27 | 27 | 0.00% |
Image 8 | 26 | 26 | 0.00% |
Image 9 | 9 | 9 | 0.00% |
Image 10 | 36 | 36 | 0.00% |
Image 11 | 28 | 28 | 0.00% |
Image 12 | 30 | 30 | 0.00% |
Image 13 | 33 | 35 | 6.06% |
Image 14 | 32 | 32 | 0.00% |
Image 15 | 31 | 31 | 0.00% |
Samples | True AP-Labeled Nanoparticle Concentration (fM) | Test AP-Labeled Nanoparticle Concentration (fM) | Relative Error |
---|---|---|---|
Image1 | 16.4222 | 16.4222 | 0.00% |
Image 2 | 9.9356 | 9.9356 | 0.00% |
Image 3 | 9.9825 | 9.9825 | 0.00% |
Image 4 | 8.7483 | 9.3610 | 7.00% |
Image 5 | 7.1905 | 7.2246 | 0.47% |
Image 6 | 13.0088 | 13.0088 | 0.00% |
Image 7 | 13.4291 | 13.4862 | 0.43% |
Image 8 | 13.5327 | 13.4730 | 0.44% |
Image 9 | 4.3976 | 4.3976 | 0.00% |
Image 10 | 11.6625 | 11.6625 | 0.00% |
Image 11 | 9.1947 | 9.1947 | 0.00% |
Image 12 | 9.6366 | 9.6366 | 0.00% |
Image 13 | 11.7421 | 12.4174 | 5.75% |
Image 14 | 9.4524 | 9.5002 | 0.51% |
Image 15 | 10.6424 | 10.6736 | 0.29% |
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Li, R.; Wang, Y.; Xu, H.; Fei, B.; Qin, B. Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy. Sensors 2017, 17, 2685. https://doi.org/10.3390/s17112685
Li R, Wang Y, Xu H, Fei B, Qin B. Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy. Sensors. 2017; 17(11):2685. https://doi.org/10.3390/s17112685
Chicago/Turabian StyleLi, Rufeng, Yibei Wang, Hong Xu, Baowei Fei, and Binjie Qin. 2017. "Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy" Sensors 17, no. 11: 2685. https://doi.org/10.3390/s17112685