Testing Criteria
Testing Criteria
Embedded Files
1. Average intensity of emitted fluorescence increases proportional to concentration %
1. Average intensity of emitted fluorescence increases proportional to concentration %
2. # of pixels processed by imager increases with concentration
2. # of pixels processed by imager increases with concentration
3. Fluorescent light intensity should decrease with time
3. Fluorescent light intensity should decrease with time
-> Values should decrease with each successive test
-> Values should decrease with each successive test
Testing Set-Up
Testing Set-Up
- The prototype was evaluated by imaging and quantifying the maximum intensity and total area of known concentrations of fluorophore outlined in the table
- The test was repeated after a week to account for the loss of inert luminescence of the fluorescent dye, resulting in a consistent decrease in max intensity and total area detected by the camera under the same parameters.
- The device was then assessed based on the following: (1) Trial 1 values were less than Trial 2, (2) the linearity of the measured values, and (3) zero offset for both intensity and area quantities. (See appendix for sample photos taken for Trial 2)
Trial 1 > Trial 2
Trial 1 > Trial 2
- Plotting both trials for total area and intensity, it can be concluded that the device is able to detect the decrease in luminescence of the fluorophore over time
Linearity and Proportionality
Linearity and Proportionality
1. Fitted Graph of the Total Area for Trial 1 and Trial 2
2. Fitted Graph of the Intensity for Trial 1 and Trial 2
Measured vs Expected Values of Total Area for Trial 1 and Trial 2
Measured vs Expected Values of Intensity for Trial 1 and Trial 2
The data taken from both trials were then fitted to a first order polynomial to analyze the linearly and offset of the results. The fitted graphs for both maximum intensity and total area of both trials are displayed above as well as tables for the expected values from the linear fit for the corresponding concentrations. Differences between the measured and expected values, normalized to the expected intensities, are also provided to quantify the linearity of the data. Most data points follow the same trendline with minimal differences, however all offset values were nonzero. This indicates that the measured intensities and areas are being imaged and quantified precisely, but there may be some errors that affect the accuracy of the results.
Conclusion
Conclusion
- The device is durable, decreasing the amount of money the hospital would need to spend on maintenance.
- Our data does show that our device and algorithm can determine a linear correlation between an increasing fluorescence intensity with the increasing concentration of the fluorophore.
- Our device, however, is unable to perform with fluorescent lifetime imaging due to the camera being too slow to capture high quality fluorescence photographs at high speed.
- Thus, our device currently can only take maximum fluorescence images to determine the amount of fluorophore in the sample.
Future Direction
Future Direction
1. Test on HIV blood samples
1. Test on HIV blood samples
-> Quantify concentration of HIV within a 5,000 copies/mL range
-> Quantify concentration of HIV within a 5,000 copies/mL range
2. Optimize camera settings and sample preparation for FLIM
2. Optimize camera settings and sample preparation for FLIM
3. Integrate different projects/components to ensure each test only costs $10-$15
3. Integrate different projects/components to ensure each test only costs $10-$15
Group 7- Low-Cost Fluorescent ImagerDepartment of BioengineeringUniversity of California, San DiegoBENG 187 - Bioengineering Design Project
Page Editor: Beatrice Tierra
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