Optimization of Single-Walled Carbon Nantube Sensor Platform for the Quantification of Extracellular Analytes

Wali Sohail Author

07/30/2021 Added

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Single Walled Carbon Nanotube sensor platform used to detect varying quantities of Nitric Oxide is optimized to have a more time efficient development process.

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[00:00:02.580]Hello, and welcome to our presentation. My name is Wally Sohail.
[00:00:05.910]And this summer I've worked with the Iris and lab for the time-based
[00:00:09.030]optimization of a single wall, carbon ENA tube,
[00:00:12.090]otherwise known as Swintt platform that will be used for the quantification of
[00:00:16.140]cellular analytes. Before I get into this,
[00:00:18.510]some relevant background information is necessary. The nitric oxide,
[00:00:22.350]and this is involved in many aspects of cell health,
[00:00:24.540]from maintenance of normal tissue homeostasis to repair of tissue injury.
[00:00:29.100]It is known that nitric oxide is essential in cell signaling,
[00:00:32.190]but it's concentration in mechanisms during healthy and disease states is
[00:00:35.580]largely unknown.
[00:00:37.830]A nanoscale real-time sensor that can quantify concentrations of extracellular
[00:00:42.300]nitric oxide in a spatial temporal fashion would allow researchers to determine
[00:00:46.710]the levels of this reactive species that is associated with specific cell types
[00:00:51.540]and functions. This would be beneficial to many areas of science,
[00:00:55.110]including understanding the basic mechanisms of cell signaling,
[00:00:58.410]determining the likelihood for increased cell flutter fibrillation and migration
[00:01:01.890]rates, and to knowing a patient's precise level of nitric oxide at the time of
[00:01:06.030]treatment, allowing for personalized drug slash dosage determination.
[00:01:11.340]Unfortunately,
[00:01:12.120]commercially available sensors cannot provide the precise spatial temporal
[00:01:15.960]quantification necessary to understand the role of nitric oxide Institute carbon
[00:01:20.910]nanotube sensors. However,
[00:01:22.140]can provide spatial temporal information about biological samples over long time
[00:01:26.760]periods due to their fluorescent recovery lack of flow photo bleaching and
[00:01:30.780]biocompatibility,
[00:01:33.300]but is there a platform with the large concentration of evenly distributed Swin
[00:01:37.050]sensors needs to be created to carry out these experiments,
[00:01:40.710]the Iris and lab previously developed this type of platform,
[00:01:43.800]but this one-time use platform takes five days to build resulting in a lot of
[00:01:47.910]wasted time between experimental planning and performance.
[00:01:51.900]This research I'll do the platform building protocol by decreasing the time for
[00:01:55.950]development while maintaining the concentration and distribution of Swin sensors
[00:02:00.420]on the platform resulting in decreased experimental downtime and increase lab
[00:02:05.190]efficiency.
[00:02:06.900]So to start off is important to understand the original method of constructing
[00:02:11.850]the Swin sensor platform, which involves five main reactions. These are the, uh,
[00:02:16.560]immersion in Pronto solution to form hydroxyl groups. Uh,
[00:02:20.490]it is then the immersion in a proxy silent or silent, silent,
[00:02:24.720]silent Istation to form a proxy groups. Uh,
[00:02:28.320]the oven curing phase for the glass slides. Uh,
[00:02:32.370]the addition of Avidan onto the surface,
[00:02:35.850]and finally the addition of biotech related swims with, uh,
[00:02:39.840]incubation period for laser analysis.
[00:02:43.650]It is sort of these core reactions,
[00:02:45.180]specifically these ones right here that the Pranam more specifically
[00:02:50.130]the Parata immersion salinization and oven curing that alternative
[00:02:54.630]alternative methods were made.
[00:02:57.120]This is because prior studies provide more evidence for the modification of the
[00:03:01.150]first three steps rather than the latter two,
[00:03:05.290]three individual experiments were conducted over the course of three weeks. Uh,
[00:03:10.120]one for the modification of each step, they are also consisted of three groups,
[00:03:14.800]each, uh, one control group, a and two alternative groups,
[00:03:18.520]B and C the control group consisted of the original
[00:03:23.140]protocol times,
[00:03:25.030]whereas the alternative groups consisted of different experimental times.
[00:03:29.200]And it's also important to note that while each of these experiments are going
[00:03:33.640]on, the rest of the protocol remained unchanged for the week.
[00:03:36.760]So only that specific step was modified for its individual experiment
[00:03:42.160]for week one, the modification, uh,
[00:03:45.430]frequent modification of the glass platform,
[00:03:47.770]immersion times in Parana solution was conducted the control group,
[00:03:52.060]a immersed the glass slides in for 14 hours. Uh,
[00:03:55.870]the control group B immerse the slides in for 45 minutes, uh,
[00:04:00.520]after pre-cleaning and NaOH and HCL,
[00:04:03.430]and the other group C immerse the slides in for one hour
[00:04:08.500]for week two, uh, the modification of the immersion times and, uh,
[00:04:13.090]of toxi silent was conducted. And then this week,
[00:04:16.390]the control group bay immersed, uh, the, uh, slides in 1%, uh,
[00:04:21.400]3g PTMS ethanol based solution for 24 hours. Uh,
[00:04:25.750]the other group be immersed, uh,
[00:04:29.080]the slides and in Tallinn based silent solution for 30 minutes.
[00:04:34.210]And the group C immersed the slides in a 10%
[00:04:38.920]cyclopentane OnBase solution for 20 minutes. Now,
[00:04:43.360]again for week three, uh,
[00:04:45.030]of in times we're testing and the control group bay, uh,
[00:04:49.090]set the slides in the oven for three hours at 150 degrees Celsius, uh,
[00:04:53.920]whereas control group or sorry, uh,
[00:04:55.830]experimental group B set the sides in the oven for one hour at 100 degrees
[00:05:00.070]Celsius a group.
[00:05:01.870]See there set the sides in the oven for one hour at 150 degrees Celsius.
[00:05:08.560]Now images were taken, uh,
[00:05:10.780]on three different points of the platform in order to accurately measure
[00:05:13.750]fluorescent spatial distribution, uh, across the glass slides.
[00:05:18.100]This was done at the end of the platform making process on the Iverson labs near
[00:05:22.510]infrared microscope that emits a laser onto the substrate in order to display
[00:05:26.800]swings, using his fluorescent properties.
[00:05:29.920]Now for image analysis prior to going into the results, um,
[00:05:34.180]the images were processed into matrices using MATLAB and then converted into
[00:05:38.440]colorized heatmaps shown right here to display and compare the mean spatial
[00:05:42.670]distribution of Swink derived fluorescent emission maps for computed by
[00:05:47.350]subtracting the average,
[00:05:48.700]some matrix of the control group bay from each individual matrix done for all
[00:05:53.330]groups in order to quantify both the fluorescent yield and spatial the
[00:05:57.830]fluorescence intensity and mean deviation RA or calculated mean
[00:06:02.570]deviation RA is given by the sum of the average
[00:06:07.010]intensity of each, uh, the entire image,
[00:06:09.920]Y subtracted from the intensity of each pixel,
[00:06:13.370]sun divided by the total number of pixels.
[00:06:17.570]Now for our results,
[00:06:19.220]all three groups came back not significantly different,
[00:06:24.680]and you could see the heat maps right here showing for each individual group
[00:06:29.780]and the mean, um, fluorescent intensity, as well as the, uh,
[00:06:34.000]fluorescent distribution shown, uh,
[00:06:38.900]the heat maps show, uh,
[00:06:41.450]even spatial distribution and the accurately depict, uh, each, um,
[00:06:47.060]uh, group.
[00:06:49.160]So for the discussion of increase in the promise solution, immersion step one,
[00:06:53.420]sanitation and the oven curing time do not significantly alter the platforms for
[00:06:58.280]us and intensity or distribution. This was analyzed with a one-way.
[00:07:01.730]I know who tests with a two key post-talk analysis for the platform.
[00:07:06.260]Immersion time for step one,
[00:07:08.120]group C is a viable alternative to the current immersion time of brute bay and
[00:07:11.990]maybe use moving forward for the silent ization process,
[00:07:16.550]the immersion in a Tallinn based a GP TMS solution for 30
[00:07:21.410]minutes, maybe using the protocol moving forward,
[00:07:23.750]as it shows the highest overall fluorescent intensity and distribution,
[00:07:29.180]the oven curing times, uh,
[00:07:30.770]there is no significant difference in fluorescent intensity and mean
[00:07:34.460]distribution. However, the P values between groups a and B,
[00:07:39.380]um,
[00:07:40.460]are higher than the p-value between groups a and C for the, uh,
[00:07:45.320]sorry, S an intensity,
[00:07:47.000]therefore option B will be used for future work
[00:07:52.910]with these three modifications.
[00:07:54.440]We decreased the time required for the construction of the wind sensor platform.
[00:07:58.490]By two days,
[00:07:59.600]making the process 40% more efficient without significantly altering the swing
[00:08:04.160]fluorescence, intensity or distribution, uh, when attached to the platform,
[00:08:08.990]this will allow for a decreased time bill spending the wind sensor platform. Uh,
[00:08:13.190]this can be visualized in these tunes here in the beginning of original protocol
[00:08:17.390]timeline. Uh, it lasted five days approximately to make these platforms,
[00:08:22.100]and now we have reduced it to around three days.
[00:08:27.260]Um, for acknowledgements,
[00:08:28.520]I'd like to thank the Iris and lab members for their support on the study.
[00:08:31.730]And this work is also supported by the national science foundation and the
[00:08:35.390]national Institute of health.
[00:08:37.250]Thank you very much for listening to my presentation and have a nice day.

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Optimization of Single-Walled Carbon Nantube Sensor Platform for the Quantification of Extracellular Analytes

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