{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/44342"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/44342","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Optimization of fluorescence lifetime imaging microscopy (FLIM) for studying the activity of enzymes in live cancer cells","abstract":"This dissertation describes the process of optimizing a Fluorescence Lifetime Imaging Microscopy (FLIM) system in order to observe the dynamics of enzymes in live cancer cells. The enzyme studied throughout this research is Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) which is a membrane-bound protein principally responsible for degrading extra-cellular matrix (ECM) proteins in the local environment of a migrating cell. However, MT1-MMP has an intricate role in the regulation of the cell’s migration separate from its simple proteolytic functions. In addition, the increased expression of MT1-MMP has been positively correlated with the invasive potential of tumor cells. In spite of the importance of MT1-MMP in understanding a cancer cell’s decision making as it leaves a tumor, very few reports have quantitatively studied the activity of this enzyme in live cells. Even fewer reports have examined the spatiotemporal activity of MT1-MMP in live cells cultured in 3-dimensional settings such as matrices of ECM proteins. These 3-dimensional settings can parallel the environment encountered by metastasizing cells in tissues. Studying live cells in 3-dimensional matrices is crucial for biologically relevant investigations. A cell’s morphology and migratory behavior can vary significantly when comparisons are made between cells cultured on two dimensional substrates and those cultured in 3-dimensional matrices. The purpose of this project was to understand the coordinated functions of MT1-MMP as live cancer cells interact with and move through a 3-dimensional matrix of ECM proteins. Specifically, we are ultimately interested in the spatiotemporal activation patterns of MT1-MMP in live cancer cells in order to build a quantitative (systems-level) model describing MT1-MMP’s role in the cell’s decision making as it is leaves a tumor site.","abstract_html":"This dissertation describes the process of optimizing a Fluorescence Lifetime Imaging Microscopy (FLIM) system in order to observe the dynamics of enzymes in live cancer cells. The enzyme studied throughout this research is Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) which is a membrane-bound protein principally responsible for degrading extra-cellular matrix (ECM) proteins in the local environment of a migrating cell. However, MT1-MMP has an intricate role in the regulation of the cell’s migration separate from its simple proteolytic functions. In addition, the increased expression of MT1-MMP has been positively correlated with the invasive potential of tumor cells. In spite of the importance of MT1-MMP in understanding a cancer cell’s decision making as it leaves a tumor, very few reports have quantitatively studied the activity of this enzyme in live cells. Even fewer reports have examined the spatiotemporal activity of MT1-MMP in live cells cultured in 3-dimensional settings such as matrices of ECM proteins. These 3-dimensional settings can parallel the environment encountered by metastasizing cells in tissues. Studying live cells in 3-dimensional matrices is crucial for biologically relevant investigations. A cell’s morphology and migratory behavior can vary significantly when comparisons are made between cells cultured on two dimensional substrates and those cultured in 3-dimensional matrices. The purpose of this project was to understand the coordinated functions of MT1-MMP as live cancer cells interact with and move through a 3-dimensional matrix of ECM proteins. Specifically, we are ultimately interested in the spatiotemporal activation patterns of MT1-MMP in live cancer cells in order to build a quantitative (systems-level) model describing MT1-MMP’s role in the cell’s decision making as it is leaves a tumor site.","abstract_has_math":false,"creators":["Eichorst, John"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biophysics & Computnl Biology","degree_department":null,"school":null,"contributors":["Wang, Yingxiao","Granick, Steve","Toussaint, Kimani C.","Gennis, Robert B."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2013,"date_issued":"2013-05-24T22:08:23Z","date_published":"2013-05-24T22:08:23Z","updated_at":"2026-07-22T22:25:34Z","subjects":["Fluorescence Lifetime Imaging Micorscopy (FLIM)","Cancer","Fluorescent Proteins","Metastasis","Enzymatic Activity"],"languages":["en"],"rights":["Copyright 2013 John Eichorst"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/44342","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Wang, Yingxiao","Granick, Steve","Toussaint, Kimani C.","Gennis, Robert B."]},{"key":"dc:creator","label":"Author","values":["Eichorst, John"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2013-05-24T22:08:23Z","2013-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biophysics & Computnl Biology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Fluorescence Lifetime Imaging Micorscopy (FLIM)","Cancer","Fluorescent Proteins","Metastasis","Enzymatic Activity"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2013 John Eichorst"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/44342"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["This dissertation describes the process of optimizing a Fluorescence Lifetime Imaging Microscopy (FLIM) system in order to observe the dynamics of enzymes in live cancer cells. 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Studying live cells in 3-dimensional matrices is crucial for biologically relevant investigations. A cell’s morphology and migratory behavior can vary significantly when comparisons are made between cells cultured on two dimensional substrates and those cultured in 3-dimensional matrices. The purpose of this project was to understand the coordinated functions of MT1-MMP as live cancer cells interact with and move through a 3-dimensional matrix of ECM proteins. 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Studying live cells in 3-dimensional matrices is crucial for biologically relevant investigations. A cell’s morphology and migratory behavior can vary significantly when comparisons are made between cells cultured on two dimensional substrates and those cultured in 3-dimensional matrices. The purpose of this project was to understand the coordinated functions of MT1-MMP as live cancer cells interact with and move through a 3-dimensional matrix of ECM proteins. 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