Introduction To facilitate measurements of chemoattractants’ effects on cancer cell migration,...
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Introduction To facilitate measurements of chemoattractants’ effects on cancer cell migration, polydimethylsiloxane (PDMS) microfluidic devices have been developed [“Study”]. These PDMS devices feature a series of two wells each connected by ten microchannels. Each microchannel is 1 mm long, 20 μm wide, and 10 μm deep. Materials and methods The experiment set-up consisted of: • Lung metastasized prostate cancer (PC3ML) cells used • Cells seeded on the wells to the left, while male rat femur bone extract was added to the wells on the right side • 5000 cells were seeded in each left well • Note: devices were sterilized before use and standard tissue culture protocols were used Acknowledgments We would like to thank NSF and UT Arlington, as well as mentors Dr. Kambiz Alavi and Mohammadreza Moghadam, for the summer research opportunity. Funding for this project was provided by the NSF (grant #EEC-1156801). Modified Data Processing Below is illustrated a flowchart of the various steps taken in order to data process with the MATLAB MeasureTool: Summary and Conclusions The MATLAB software features a number of advantages over the original AxioVision software section. The program can be run on any platform, and as a MATLAB code is open to future additions and modifications if the need arises. The modified data processing also includes a cell counting function, which originally had to be computed and entered by hand. Finally, the program makes steps towards minimizing the amount of manual entry necessary in data processing, saving tedious time spent in the original process. When weighing the effectiveness of the MeasureTool with the AxioVision measuring tool, a random sampling process was conducted to gage the differences in how the two different measuring tools compared. This process led to the calculation of low percentage errors, indicating the MeasureTool was just as accurate in measuring and converting the pixels to microns (μm). As an outside source was consulted, however, to obtain the comparison measurements in AxioVision, the process is still open to differences. Since the measurements and counting depend on interpretation, there are some inherent discrepancies. Increasing the number of samples and having several independent assessments could help minimize errors. Future work, therefore, would include attempts to further decrease the amount of manual work required in data processing. Such improvements would include recording data from multiple pictures organized into multiple tables in one text file. This would reduce the amount of manual transferring to Excel that is needed, and improve the overall efficiency of the process to a greater degree. Arshia Ali , Akash Sharma, Smitha Rao, J. -C. Chiao Electrical Engineering Summer REU: Sensors, The University of Texas at Arlington, Arlington, Texas 76019 Literature cited “Study of Prostate Cancer Cell Line PC3-ML Chemotaxsis to Epidermal Growth Factor with a BIOMEMS Device,” U. Tata, S M.N. Rao, A. Sharma, K. Pabba, K. Pokhrel, V. K. Lin, H. Cao, and J.-C. Chiao, The 3 rd International Workshop on Nanotechnology and Application, IWNA2011, Vung Tau Vietnam, Nov. 10-12 2011. Figure 1. Each PDMS device consists of multiple device systems, each of which feature two wells connected by microchannels. Figure 2. In some cases, nanoparticles were added to the cells seeded on the left hand side. Three different types of nanoparticles were used: gold (A), iron oxide (B), and nickel (C). Iron oxide nanoparticles were typically used for their magnetic properties and minimal cytotoxity. Figure 3. After running the MATLAB program, the following MeasureTool will pop up. To begin editing a picture, click on the Browse button and find the appropriate image. For further information Please contact [email protected] (Arshia Ali) or [email protected] (Dr. Smitha Rao). More information on this project and related projects can be obtained at http://www.uta.edu/faculty/jcchiao/research_front .htm. Data Processing Cancer Cell Migration in Microfluidic Devices Abstract Understanding why cancer cells metastasize would be influential on the development of cancer treatments. The chemical effects of multiple male rat organs extracts were measured on the movement of prostate cancer cells (PC3ML). The need for qualitative data in measuring the chemoattractant’s effects led to the development of PDMS microfluidic devices that can measure cell migration. These devices are characterized by two wells connected by 10 microchannels. Experiment set-up consisted of cells seeded on the left side wells of the device, and chemoattractants added to the right. The method for analyzing this qualitative method had issues as it required manual cell counting and manual data entry from the imaging measurement system into an Excel file to be processed. In an effort to improve efficiency, a MATLAB code was written to complete three main functions: performing cell count, measuring cell migration, and exporting data to a text file. . Results Figure 4. Once the image file has been selected, the channel lengths need to be calibrated from pixels to microns. Each of the ten microchannel is 1000 μm (1 mm) long. Figure 5. After calibrating the image, migration measurements can be made. Select the Distance button to do so, and double click the line to confirm a measurement. If an error is made while measuring, the Edit and Delete buttons allow choosing which measurements to modify. Figure 6. Once an image has been modified to include migration measurements, the distances can be saved to a text file. Click on the Text button and name the file. MeasureTool AxioVision Figure 7. The migration of the PC3ML cells was processed over 5 days. The control sample shows minimal overall migration. Meanwhile, the femur extract samples of concentrations 2μg and 8μg demonstrated downwards trends in overall cell migration before increasing once more. As for the 4μg femur extract sample, no migration was recorded until the fifth and final day of the experiment. Figure 8. For this example image, a percentage error of 1.314% was calculated for the two comparing measurements in the second microchannel. This low percentage error demonstrates that MeasureTool is comparable with AxioVision as a cell migration measurement tool. As displayed, however, there are still noticeable differences open to any measurement method, as determining cell count and cell migration is still determined by human judgment. In the above example, the MeasureTool user perceived a cell in the seventh microchannel while the AxioVision user did not.
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Transcript of Introduction To facilitate measurements of chemoattractants’ effects on cancer cell migration,...
IntroductionTo facilitate measurements of chemoattractants’ effects on cancer cell migration, polydimethylsiloxane (PDMS) microfluidic devices have been developed [“Study”]. These PDMS devices feature a series of two wells each connected by ten microchannels. Each microchannel is 1 mm long, 20 μm wide, and 10 μm deep.
Materials and methodsThe experiment set-up consisted of: • Lung metastasized prostate cancer (PC3ML) cells used• Cells seeded on the wells to the left, while male rat femur
bone extract was added to the wells on the right side • 5000 cells were seeded in each left well• Note: devices were sterilized before use and standard tissue
culture protocols were used AcknowledgmentsWe would like to thank NSF and UT Arlington, as well as mentors Dr. Kambiz Alavi and Mohammadreza Moghadam, for the summer research opportunity. Funding for this project was provided by the NSF (grant #EEC-1156801).
Modified Data Processing Below is illustrated a flowchart of the various steps taken in order to data process with the MATLAB MeasureTool:
Summary and ConclusionsThe MATLAB software features a number of advantages over the original AxioVision software section. The program can be run on any platform, and as a MATLAB code is open to future additions and modifications if the need arises. The modified data processing also includes a cell counting function, which originally had to be computed and entered by hand. Finally, the program makes steps towards minimizing the amount of manual entry necessary in data processing, saving tedious time spent in the original process.
When weighing the effectiveness of the MeasureTool with the AxioVision measuring tool, a random sampling process was conducted to gage the differences in how the two different measuring tools compared. This process led to the calculation of low percentage errors, indicating the MeasureTool was just as accurate in measuring and converting the pixels to microns (μm). As an outside source was consulted, however, to obtain the comparison measurements in AxioVision, the process is still open to differences. Since the measurements and counting depend on interpretation, there are some inherent discrepancies. Increasing the number of samples and having several independent assessments could help minimize errors.
Future work, therefore, would include attempts to further decrease the amount of manual work required in data processing. Such improvements would include recording data from multiple pictures organized into multiple tables in one text file. This would reduce the amount of manual transferring to Excel that is needed, and improve the overall efficiency of the process to a greater degree.
Arshia Ali, Akash Sharma, Smitha Rao, J. -C. Chiao
Electrical Engineering Summer REU: Sensors, The University of Texas at Arlington, Arlington, Texas 76019
Literature cited“Study of Prostate Cancer Cell Line PC3-ML Chemotaxsis to
Epidermal Growth Factor with a BIOMEMS Device,” U. Tata, S M.N. Rao, A. Sharma, K. Pabba, K. Pokhrel, V. K. Lin, H. Cao, and J.-C. Chiao, The 3rd International Workshop on Nanotechnology and Application, IWNA2011, Vung Tau Vietnam, Nov. 10-12 2011.
Figure 1. Each PDMS device consists of multiple device systems, each of which feature two wells connected by microchannels.
Figure 2. In some cases, nanoparticles were added to the cells seeded on the left hand side. Three different types of nanoparticles were used: gold (A), iron oxide (B), and nickel (C). Iron oxide nanoparticles were typically used for their magnetic properties and minimal cytotoxity.
Figure 3. After running the MATLAB program, the following MeasureTool will pop up. To begin editing a picture, click on the Browse button and find the appropriate image.
For further informationPlease contact [email protected] (Arshia Ali) or [email protected] (Dr. Smitha Rao).
More information on this project and related projects can be obtained at http://www.uta.edu/faculty/jcchiao/research_front.htm.
Data Processing Cancer Cell Migration in Microfluidic Devices
AbstractUnderstanding why cancer cells metastasize would be influential on the development of cancer treatments. The chemical effects of multiple male rat organs extracts were measured on the movement of prostate cancer cells (PC3ML). The need for qualitative data in measuring the chemoattractant’s effects led to the development of PDMS microfluidic devices that can measure cell migration. These devices are characterized by two wells connected by 10 microchannels. Experiment set-up consisted of cells seeded on the left side wells of the device, and chemoattractants added to the right. The method for analyzing this qualitative method had issues as it required manual cell counting and manual data entry from the imaging measurement system into an Excel file to be processed. In an effort to improve efficiency, a MATLAB code was written to complete three main functions: performing cell count, measuring cell migration, and exporting data to a text file.
.
Results
Figure 4. Once the image file has been selected, the channel lengths need to be calibrated from pixels to microns. Each of the ten microchannel is 1000 μm (1 mm) long.
Figure 5. After calibrating the image, migration measurements can be made. Select the Distance button to do so, and double click the line to confirm a measurement. If an error is made while measuring, the Edit and Delete buttons allow choosing which measurements to modify.
Figure 6. Once an image has been modified to include migration measurements, the distances can be saved to a text file. Click on the Text button and name the file.
MeasureTool AxioVision
Figure 7. The migration of the PC3ML cells was processed over 5 days. The control sample shows minimal overall migration. Meanwhile, the femur extract samples of concentrations 2μg and 8μg demonstrated downwards trends in overall cell migration before increasing once more. As for the 4μg femur extract sample, no migration was recorded until the fifth and final day of the experiment.
Figure 8. For this example image, a percentage error of 1.314% was calculated for the two comparing measurements in the second microchannel. This low percentage error demonstrates that MeasureTool is comparable with AxioVision as a cell migration measurement tool. As displayed, however, there are still noticeable differences open to any measurement method, as determining cell count and cell migration is still determined by human judgment. In the above example, the MeasureTool user perceived a cell in the seventh microchannel while the AxioVision user did not.
