2026-09-19T06:28:42Zhttps://dspace.mit.edu/server/oai/requestoai:dspace.mit.edu:1721.1/404772022-01-13T07:54:36Zcom_1721.1_7582com_1721.1_7581col_1721.1_131024
Paul T. Matsudaira.
Reed, Stephanie M
Massachusetts Institute of Technology. Dept. of Mechanical Engineering.
Massachusetts Institute of Technology. Department of Mechanical Engineering
2008-02-27T22:29:58Z
2008-02-27T22:29:58Z
2007
2007
http://hdl.handle.net/1721.1/40477
191750168
Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007.
Includes bibliographical references.
Cytoskeletal proteins function as dynamic, complex components involved in cellular structure and signaling. Characterizing the roles of such proteins would greatly benefit many research areas, including the study of cancer and protein-related diseases. There is currently no accurate, high throughput method of image analysis that clearly describes protein behavior within the cell. In addressing this problem, we chose to characterize proteins based on the colocalization parameter-the amount of overlap between two objects or signals. We aimed to create a single parameter that quantitatively defined colocalization yet complemented biological intuition about a complicated system. Cell culture techniques were used to transfect HeLa cells with four "marker" GFP-tagged protein constructs. Cells were fluorescently labeled in three channels-Hoechst for nucleus, Texas Red phalloidin for actin, and GFP for protein-and images were captured using Cellomics scanning microscopy. After collecting data and testing software applications, we analyzed our data with Definiens software and developed a flexible, comprehensible method of quantifying colocalization using minimal parameters.
by Stephanie M. Reed.
S.B.
35 p.
eng
Massachusetts Institute of Technology
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
Mechanical Engineering.
Analyzing GFP-tagged cytoskeletal protein colocalization in human carcinoma cells
Thesis
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