Home
Resource
Frequency-domain fluorescence lifetime imaging microscopy for metabolic imaging of autofluorescent coenzymes NADH and FAD in biofilms
PDF
PDF format is widely accepted and good for printing.
Plug-in required
PDF-1(6.53 MB)
Citation & Export
View Usage Statistics
Staff View

Citation & Export
Hide

Simple citation

Chakraborty, Anushka. Frequency-domain fluorescence lifetime imaging microscopy for metabolic imaging of autofluorescent coenzymes NADH and FAD in biofilms. Retrieved from https://doi.org/doi:10.7282/t3-h9nm-vc57

Export

Click here for information about Citation Management Tools at Rutgers.

Statistics
Hide

Description

TitleFrequency-domain fluorescence lifetime imaging microscopy for metabolic imaging of autofluorescent coenzymes NADH and FAD in biofilms
NameChakraborty, Anushka (author); Pierce, Mark (chair); Boustany, Nada (member); Shrirao, Anil (member); Rutgers University; School of Graduate Studies
Date Created2023
Other Date2023-01 (degree)
SubjectBiomedical engineering, Fluorescence microscopy
Extent89 pages : illustrations
DescriptionFluorescence lifetime imaging microscopy (FLIM) is an imaging technique that canmeasure fluorophore lifetime in biological cells and tissues. FLIM can provide additional information beyond conventional fluorescence intensity or spectral imaging through knowledge of the fluorescence lifetime of a sample, which is an intrinsic trait of a fluorophore that can be extremely sensitive to the surrounding environment of the fluorophore, including factors such as pH, temperature, oxygenation, viscosity, and binding state.
In this thesis, a prototype frequency-domain FLIM system was evaluated for its ability to measure fluorescence lifetimes of the autofluorescent coenzymes NADH and FAD in biological samples. To produce the most accurate, reliable, and stable fluorescence lifetime measurements, calibration methods using fluorophores with known lifetimes were established, as well as experiments that determined the best combination of parameters (image size (number of pixels), scan size (field-of-view), integration time, and PMT gain) to acquire weak signals on the new FLIM system. To assess whether this FLIM system could image the lifetimes of NADH and FAD within biofilms, FLIM was performed on oral biofilms prepared from human saliva on hydroxyapatite discs. Fluorescence lifetimes in untreated control groups were compared to treatment groups that received anti-bacterial mouthwash.
After quantitative analysis was performed on images of control and treatment biofilms, the mouthwash treatment was found to have an effect on the fluorescence lifetimes, as the mean fluorescence lifetime of the control biofilms differed from the treatment biofilms. The change in fluorescence lifetime measured in NADH was more significant than it was for FAD. Some of the results from the quantitative analysis of the experiments ran were that the NADH lifetime for the control biofilms was calculated to be 1.41± 0.04 ns and the mean FAD lifetime was 1.39± 0.03 ns over three samples imaged at one session.
This thesis describes instrument calibration, image acquisition, and image analysis methods for measuring NADH and FAD autofluorescence in biofilms. These studies provide support for further development of FLIM as a tool that can be used in multiple health-related applications for non-invasive imaging and detection of alterations in metabolism.
NoteM.S.
NoteIncludes bibliographical references
Genretheses
Persistent URLhttps://doi.org/doi:10.7282/t3-h9nm-vc57
LanguageEnglish
CollectionSchool of Graduate Studies Electronic Theses and Dissertations
Organization NameRutgers, The State University of New Jersey
RightsThe author owns the copyright to this work.
Version 8.5.5
Rutgers University Libraries - Copyright ©2024