Lab #11 Report

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447

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Mechanical Engineering

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Jan 9, 2024

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McKenna Nichols BME 447 Loeffler 29 November 2023 Chapter 14 Lab Report: Lab-on-a-Chip Biosensors Results Task 4: Bradford Assay with Paper-Based LOC Without Flash With Flash BSA Concentration (mg/mL) 0 1 2 0 1 2 Red Intensity (I) 153.632 148.262 132.1 158.259 136.259 127.846 Green Intensity (I) 167.512 178.734 160.317 185.254 194.178 178.436 Blue Intensity 160 189.748 167.885 191.433 231.153 213.818 [red] 𝐴 = 𝑙𝑜𝑔( 𝐼 0 𝐼 ) 0 0.0154518 0.0655789 0 0.0628096 0.0926813 [green] 𝐴 = 𝑙𝑜𝑔( 𝐼 0 𝐼 ) 0 -0.0281612 0.0190663 0 -0.0204325 0.0162851

Further Study: LAMP in LOC Time (min) Blue Intensity Red Intensity Green Intensity Green/Red 0 173.235 205.903 183.582 0.89159 5 184.797 221.602 203.590 0.91872 10 178.814 224.943 205.981 0.91570 15 177.251 221.686 205.449 0.92676 20 180.083 226.320 207.611 0.91733 25 185.472 231.214 213.528 0.92351 30 178.382 228.570 219.284 0.95937 35 170.709 223.642 208.938 0.93425 40 168.302 222.529 210.308 0.94508

Discussion In the first task of this lab, we began by placing 3 paper dots in a plastic dish. For each solution, 2 mg/mL, 1 mg/mL, and 0 mg/ml , we added 2 sample onto the center of each dot. µ𝐿 After letting the paper dots dry for 5 minutes, we added 8 of Bradford Reagent to the center µ𝐿 of each paper dot and then again let the paper dots dry for 5 minutes. We then took two pictures of each paper dot, one with flash and one without flash, at a constant distance (between 4 and 6 inches). Then using the ImageJ programmer, we obtained the mean intensity values for red, green, and blue. From here we calculated the absorbances for the red and green intensities. We then graphed the absorbance versus the concentration of BSA. Based on our results we see that the absorbance for the red intensities increases at a faster rate than the absorbances of the green intensities.

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In the further study portion of this lab, we began by vortexing all reagents. Then in a tube, we added 6.25 LAMP master mix, 1.25 primer mix, 0.5 target, and 4.5 µ𝐿 µ𝐿 µ𝐿 µ𝐿 nuclease free water. Again, we vortexed to mix reagents. We then placed a glass slide on a hot plate followed by a paper dot. We dispensed 10 of LAMP mixture onto the paper dot. We µ𝐿 then added 5 of mineral oil to prevent evaporation and then sealed it with a cover slip. We left µ𝐿 the paper dot on the hot plate for a total of 40 minutes taking a picture every 5 minutes. Then using the ImageJ programmer, we obtained the mean intensity values for the red, green, and blue channels. We calculated the Green/Red ratio and then plotted it versus time. Our results showed an overall increase in the green to red ratio as time increased. Review Questions 14.4 Describe the linearity ( ) of your standard curves with versus without using flash. If nonlinear, 𝑅 2 briefly explain why. Both the standard curves for the Green wavelength, with and without using flash, are nonlinear. This is because of their values. The value for the without flash standard curve is 0.161 and 𝑅 2 𝑅 2 the value for the flash standard curve is 0.062. Both the standard curves for the Red 𝑅 2 wavelength, with and without using flash, are linear. This is because of their values. The 𝑅 2 𝑅 2 value for the without flash standard curve is 0.915 and the value for the flash standard curve 𝑅 2 is 0.96. The green may be nonlinear because 14.5 Describe the sensitivity (slope) of your standard curves with red versus green pixel intensities. Both the slopes on for the Red intensities are much greater. The values on the red graph increase much quicker than those on the green graph. All the slopes are positive, meaning that as the

concentration of the BSA increases, the absorbance also increases. The green pixel intensities do increase absorbance at a much slower rate than the red. Extra Why are LOC devices so useful? What kinds of settings can they be applied in? LOC devices are so useful because they have a low consumption of chemicals, they allow for rapid analysis, and they can be produced and sold at a much lower cost than other types of devices. These devices are capable of integrating several laboratory functions onto a small surface area. LOC devices could easily be applied to detect diseases in third world countries. Another application is using LOCs for chemical analysis such as mass spectrometry.