{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/105131"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/105131","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Label-free multiphoton microscopy for imaging transient metabolic dynamics in living cells and tissue","abstract":"Cellular metabolism plays a critical role in human health and homeostasis and is implicated in a large number of pathological conditions. While clinical imaging tools have emerged to probe metabolism at the tissue and organ level, the tools to probe metabolic dynamics at the cellular level have been slow to develop. This thesis represents a step toward realizing one such tool through the use of two-photon fluorescence lifetime imaging microscopy (2P-FLIM) of reduced nicotinamide adenine dinucleotide (NADH). This autofluorescent co-enzyme is involved in both aerobic and anaerobic metabolic processes which can be differentiated utilizing this advanced imaging approach. This thesis first presents a study of cell death dynamics in vivo, with cellular resolution, with a custom-built microscope utilizing a commercial 2P-FLIM detection system. Motivated by the limitations of this study in observing the early dynamics, a high-speed 2P-FLIM instrument is developed and characterized. This developed system is then directly applied to study the rapid, transient metabolic dynamics of cell death, providing new insight into this dynamic metabolic environment. Finally, this tool is combined with fluorescence calcium imaging to study the metabolic dynamics in neuronal activation, revealing a strong cell-specific response to brain activity in dissociated hippocampal cultures. These studies together demonstrate the potential of dynamic metabolic imaging as a tool for both basic scientific research and potential clinical translation. Through further development of these imaging approaches, the complex relationship between cellular metabolism and human health and disease can be further disentangled, providing potential benefits for both future biomedical research and clinical outcomes.","abstract_html":"Cellular metabolism plays a critical role in human health and homeostasis and is implicated in a large number of pathological conditions. While clinical imaging tools have emerged to probe metabolism at the tissue and organ level, the tools to probe metabolic dynamics at the cellular level have been slow to develop. This thesis represents a step toward realizing one such tool through the use of two-photon fluorescence lifetime imaging microscopy (2P-FLIM) of reduced nicotinamide adenine dinucleotide (NADH). This autofluorescent co-enzyme is involved in both aerobic and anaerobic metabolic processes which can be differentiated utilizing this advanced imaging approach. This thesis first presents a study of cell death dynamics in vivo, with cellular resolution, with a custom-built microscope utilizing a commercial 2P-FLIM detection system. Motivated by the limitations of this study in observing the early dynamics, a high-speed 2P-FLIM instrument is developed and characterized. This developed system is then directly applied to study the rapid, transient metabolic dynamics of cell death, providing new insight into this dynamic metabolic environment. Finally, this tool is combined with fluorescence calcium imaging to study the metabolic dynamics in neuronal activation, revealing a strong cell-specific response to brain activity in dissociated hippocampal cultures. These studies together demonstrate the potential of dynamic metabolic imaging as a tool for both basic scientific research and potential clinical translation. Through further development of these imaging approaches, the complex relationship between cellular metabolism and human health and disease can be further disentangled, providing potential benefits for both future biomedical research and clinical outcomes.","abstract_has_math":false,"creators":["Bower, Andrew John"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Electrical & Computer Engr","degree_department":null,"school":null,"contributors":["Boppart, Stephen A.","Gao, Liang","Gillette, Martha U.","Popescu, Gabriel"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-08-23T20:44:30Z","date_published":"2019-08-23T20:44:30Z","updated_at":"2026-07-22T22:24:44Z","subjects":["multiphoton microscopy","fluorescence lifetime imaging microscopy","optical metabolic imaging","high-speed microscopy"],"languages":["en"],"rights":["Copyright 2019 Andrew Bower"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/105131","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Boppart, Stephen A.","Gao, Liang","Gillette, Martha U.","Popescu, Gabriel"]},{"key":"dc:creator","label":"Author","values":["Bower, Andrew John"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-08-23T20:44:30Z","2021-08-24T09:15:31Z","2019-02-11","2019-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Electrical & Computer Engr"]},{"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":["multiphoton microscopy","fluorescence lifetime imaging microscopy","optical metabolic imaging","high-speed microscopy"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2019 Andrew Bower"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/105131"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Cellular metabolism plays a critical role in human health and homeostasis and is implicated in a large number of pathological conditions. While clinical imaging tools have emerged to probe metabolism at the tissue and organ level, the tools to probe metabolic dynamics at the cellular level have been slow to develop. This thesis represents a step toward realizing one such tool through the use of two-photon fluorescence lifetime imaging microscopy (2P-FLIM) of reduced nicotinamide adenine dinucleotide (NADH). This autofluorescent co-enzyme is involved in both aerobic and anaerobic metabolic processes which can be differentiated utilizing this advanced imaging approach. This thesis first presents a study of cell death dynamics in vivo, with cellular resolution, with a custom-built microscope utilizing a commercial 2P-FLIM detection system. Motivated by the limitations of this study in observing the early dynamics, a high-speed 2P-FLIM instrument is developed and characterized. This developed system is then directly applied to study the rapid, transient metabolic dynamics of cell death, providing new insight into this dynamic metabolic environment. Finally, this tool is combined with fluorescence calcium imaging to study the metabolic dynamics in neuronal activation, revealing a strong cell-specific response to brain activity in dissociated hippocampal cultures. These studies together demonstrate the potential of dynamic metabolic imaging as a tool for both basic scientific research and potential clinical translation. Through further development of these imaging approaches, the complex relationship between cellular metabolism and human health and disease can be further disentangled, providing potential benefits for both future biomedical research and clinical outcomes.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-05-01","The student, Andrew Bower, accepted the attached license on 2019-02-10 at 12:42.","The student, Andrew Bower, submitted this Dissertation for approval on 2019-02-10 at 12:47.","This Dissertation was approved for publication on 2019-02-11 at 11:50.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13381 on 2019-08-22 at 16:19:47","Made available in DSpace on 2019-08-23T20:44:30Z (GMT). No. of bitstreams: 12 BOWER-DISSERTATION-2019.pdf: 5455650 bytes, checksum: 6e9bac95df47407abd127291a6d3a00b (MD5) Chapter 3.pdf: 196487 bytes, checksum: 4aea9cbae4c24ec67339cb07ee1e1c8a (MD5) Chapter 4.pdf: 102278 bytes, checksum: 9c08fe89c1d602481f6f8e7f92b1bc15 (MD5) Figure 1.1.pdf: 133667 bytes, checksum: baf0fd8f3215509ac6b3d4ba36a2ec1d (MD5) Figure 2.1.pdf: 267928 bytes, checksum: c4b08f63561882c10e25c53a567aecd3 (MD5) Figure 2.12.pdf: 654544 bytes, checksum: 2621b7b0de96eba06a438114dff76068 (MD5) Figure 2.13.pdf: 188032 bytes, checksum: aecab0fca0300c5fff090f0b2580397c (MD5) Figure 2.14.pdf: 1555255 bytes, checksum: 314af75274a1d9eb88de6e9e331d5ae6 (MD5) Figure 2.2.pdf: 128164 bytes, checksum: ac069f9aea780529e5a829f858cd24c3 (MD5) Figure 2.3.pdf: 122492 bytes, checksum: 1d1af057e626ee10413281d74851b6d1 (MD5) Figure 6.1.pdf: 122121 bytes, checksum: 5add4dacf74e2e24a2aad4a559eff3d8 (MD5) LICENSE.txt: 4209 bytes, checksum: 270fa8e4a328f4442a2242ffd1fcc5d9 (MD5) Previous issue date: 2019-02-11","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:44:50Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:46:41Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:47:38Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:48:32Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 112250 on 2021-08-24T09:15:31Z."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Label-free multiphoton microscopy for imaging transient metabolic dynamics in living cells and tissue"]}]}],"canonical_facts":{"dc:contributor":["Boppart, Stephen A.","Gao, Liang","Gillette, Martha U.","Popescu, Gabriel"],"dc:creator":["Bower, Andrew John"],"dc:date":["2019-08-23T20:44:30Z","2021-08-24T09:15:31Z","2019-02-11","2019-05"],"dc:description":["Cellular metabolism plays a critical role in human health and homeostasis and is implicated in a large number of pathological conditions. While clinical imaging tools have emerged to probe metabolism at the tissue and organ level, the tools to probe metabolic dynamics at the cellular level have been slow to develop. This thesis represents a step toward realizing one such tool through the use of two-photon fluorescence lifetime imaging microscopy (2P-FLIM) of reduced nicotinamide adenine dinucleotide (NADH). This autofluorescent co-enzyme is involved in both aerobic and anaerobic metabolic processes which can be differentiated utilizing this advanced imaging approach. This thesis first presents a study of cell death dynamics in vivo, with cellular resolution, with a custom-built microscope utilizing a commercial 2P-FLIM detection system. Motivated by the limitations of this study in observing the early dynamics, a high-speed 2P-FLIM instrument is developed and characterized. This developed system is then directly applied to study the rapid, transient metabolic dynamics of cell death, providing new insight into this dynamic metabolic environment. Finally, this tool is combined with fluorescence calcium imaging to study the metabolic dynamics in neuronal activation, revealing a strong cell-specific response to brain activity in dissociated hippocampal cultures. These studies together demonstrate the potential of dynamic metabolic imaging as a tool for both basic scientific research and potential clinical translation. Through further development of these imaging approaches, the complex relationship between cellular metabolism and human health and disease can be further disentangled, providing potential benefits for both future biomedical research and clinical outcomes.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-05-01","The student, Andrew Bower, accepted the attached license on 2019-02-10 at 12:42.","The student, Andrew Bower, submitted this Dissertation for approval on 2019-02-10 at 12:47.","This Dissertation was approved for publication on 2019-02-11 at 11:50.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13381 on 2019-08-22 at 16:19:47","Made available in DSpace on 2019-08-23T20:44:30Z (GMT). No. of bitstreams: 12 BOWER-DISSERTATION-2019.pdf: 5455650 bytes, checksum: 6e9bac95df47407abd127291a6d3a00b (MD5) Chapter 3.pdf: 196487 bytes, checksum: 4aea9cbae4c24ec67339cb07ee1e1c8a (MD5) Chapter 4.pdf: 102278 bytes, checksum: 9c08fe89c1d602481f6f8e7f92b1bc15 (MD5) Figure 1.1.pdf: 133667 bytes, checksum: baf0fd8f3215509ac6b3d4ba36a2ec1d (MD5) Figure 2.1.pdf: 267928 bytes, checksum: c4b08f63561882c10e25c53a567aecd3 (MD5) Figure 2.12.pdf: 654544 bytes, checksum: 2621b7b0de96eba06a438114dff76068 (MD5) Figure 2.13.pdf: 188032 bytes, checksum: aecab0fca0300c5fff090f0b2580397c (MD5) Figure 2.14.pdf: 1555255 bytes, checksum: 314af75274a1d9eb88de6e9e331d5ae6 (MD5) Figure 2.2.pdf: 128164 bytes, checksum: ac069f9aea780529e5a829f858cd24c3 (MD5) Figure 2.3.pdf: 122492 bytes, checksum: 1d1af057e626ee10413281d74851b6d1 (MD5) Figure 6.1.pdf: 122121 bytes, checksum: 5add4dacf74e2e24a2aad4a559eff3d8 (MD5) LICENSE.txt: 4209 bytes, checksum: 270fa8e4a328f4442a2242ffd1fcc5d9 (MD5) Previous issue date: 2019-02-11","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:44:50Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:46:41Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:47:38Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112250 Lift date: 2021-08-23T20:48:32Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 112250 on 2021-08-24T09:15:31Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/105131"],"dc:language":["en"],"dc:rights":["Copyright 2019 Andrew Bower"],"dc:subject":["multiphoton microscopy","fluorescence lifetime imaging microscopy","optical metabolic imaging","high-speed microscopy"],"dc:title":["Label-free multiphoton microscopy for imaging transient metabolic dynamics in living cells and tissue"],"dc:type":["text"],"thesis:degree_discipline":["Electrical & Computer Engr"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:24:44Z"}