A cell-based sensor of fluid shear stress for microfluidics
Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2013.
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| Format: | Thesis |
| Language: | eng |
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Massachusetts Institute of Technology
2013
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| Online Access: | http://hdl.handle.net/1721.1/82372 |
| _version_ | 1826210208519553024 |
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| author | Varma, Sarvesh |
| author2 | Joel Voldman. |
| author_facet | Joel Voldman. Varma, Sarvesh |
| author_sort | Varma, Sarvesh |
| collection | MIT |
| description | Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2013. |
| first_indexed | 2024-09-23T14:46:10Z |
| format | Thesis |
| id | mit-1721.1/82372 |
| institution | Massachusetts Institute of Technology |
| language | eng |
| last_indexed | 2024-09-23T14:46:10Z |
| publishDate | 2013 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/823722019-04-11T03:27:44Z A cell-based sensor of fluid shear stress for microfluidics Cell-based sensor of FSS for microfluidics Varma, Sarvesh Joel Voldman. Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science. Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science. Electrical Engineering and Computer Science. Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2013. Cataloged from PDF version of thesis. Includes bibliographical references (p. 100-104). Fluid flow is an essential feature of every microsystem involving cell handling, culture or sorting. The particular application determines the relevant flow rates used in a device. Flows inevitably generate fluid shear stress (FSS) that may cause undesirable physiological cell stress. Simple assays of cell viability, morphology or growth are typically reported to indicate any gross disturbances to cell physiology. However, no straightforward metric exists to specifically evaluate physiological implications of FSS within microfluidic devices, or among competing microfluidic technologies).This thesis presents the first genetically encoded cell sensors that fluoresce in a quantitative fashion upon FSS pathway activation. A transcriptional sensor was chosen, meaning that fluorescence would be turned on when transcription of a relevant protein was initiated. Creating an effective transcriptional sensor requires identifying an appropriate inducible promoter to drive fluorescence expression upon FSS. To this end, the native mechanotransduction of a widely used and easy-to-culture cell line (NIH3T3s) was elucidated by culturing them in a microfluidic device and applying logarithmic FSS via microfluidic perfusion. A panel of shear-responsive genes was analyzed using qRT-PCR, which resulted in the choice of EGR-1 upregulation as the node for FSS detection. A reporter plasmid with a minimal EGR-1 promoter driving the expression of Turbo-RFP fluorescence was chosen and the cell sensor was created by stable transfection and clonal selection. Inducing the pathway with Phorbol-myristate-acetate resulted in fluorescence induction by both microscopy and flow cytometry, verifying the sensor functionality. The fluorescence activation was then characterized across PMA doses and durations. Next, the sensors were tested using multiple duration microfluidic perfusions, where it was noted that the mean induced fluorescence intensity correlated to applied FSS intensity, as desired. It is anticipated that these cell sensors will have wide application in the microsystems community, allowing the device designer to engineer systems with acceptable FSS, and allowing the end-user to evaluate impact of FSS upon their assay of interest. by Sarvesh Varma. S.M. 2013-11-18T19:15:03Z 2013-11-18T19:15:03Z 2013 2013 Thesis http://hdl.handle.net/1721.1/82372 862074616 eng M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 104 p. application/pdf Massachusetts Institute of Technology |
| spellingShingle | Electrical Engineering and Computer Science. Varma, Sarvesh A cell-based sensor of fluid shear stress for microfluidics |
| title | A cell-based sensor of fluid shear stress for microfluidics |
| title_full | A cell-based sensor of fluid shear stress for microfluidics |
| title_fullStr | A cell-based sensor of fluid shear stress for microfluidics |
| title_full_unstemmed | A cell-based sensor of fluid shear stress for microfluidics |
| title_short | A cell-based sensor of fluid shear stress for microfluidics |
| title_sort | cell based sensor of fluid shear stress for microfluidics |
| topic | Electrical Engineering and Computer Science. |
| url | http://hdl.handle.net/1721.1/82372 |
| work_keys_str_mv | AT varmasarvesh acellbasedsensoroffluidshearstressformicrofluidics AT varmasarvesh cellbasedsensoroffssformicrofluidics AT varmasarvesh cellbasedsensoroffluidshearstressformicrofluidics |