Deconvolution of fluorescence lifetime imaging microscopy by a library of exponentials

Campos-Delgado, Daniel U.; Navarro, Omar Gutiérrez; Arce-Santana, Edgar R.; Walsh, Alex J.; Skala, Melissa C.; Jo, Javier A.

doi:10.1364/oe.23.023748

Cited by 15 publications

(16 citation statements)

References 26 publications

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“…Deconvolution includes variations on the least squares approach discussed above, stretched exponential/lifetime moment analysis, and methods of transformation (e.g., Fourier and Laplace transforms). 177,179,[194][195][196][197][198] Stretched exponential and lifetime moment methods are fundamentally similar because the lifetimes of individual species are estimated from the total measured decay distribution. 177,179 Stretched exponential and lifetime moment methods have specific implementations for time-domain or frequency-domain data.…”

Section: Deconvolution Analysismentioning

confidence: 99%

Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications

et al. 2020

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Significance: Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to distinguish the unique molecular environment of fluorophores. FLIM measures the time a fluorophore remains in an excited state before emitting a photon, and detects molecular variations of fluorophores that are not apparent with spectral techniques alone. FLIM is sensitive to multiple biomedical processes including disease progression and drug efficacy. Aim: We provide an overview of FLIM principles, instrumentation, and analysis while highlighting the latest developments and biological applications. Approach: This review covers FLIM principles and theory, including advantages over intensitybased fluorescence measurements. Fundamentals of FLIM instrumentation in time-and frequencydomains are summarized, along with recent developments. Image segmentation and analysis strategies that quantify spatial and molecular features of cellular heterogeneity are reviewed. Finally, representative applications are provided including high-resolution FLIM of cell-and organelle-level molecular changes, use of exogenous and endogenous fluorophores, and imaging protein-protein interactions with Förster resonance energy transfer (FRET). Advantages and limitations of FLIM are also discussed. Conclusions: FLIM is advantageous for probing molecular environments of fluorophores to inform on fluorophore behavior that cannot be elucidated with intensity measurements alone. Development of FLIM technologies, analysis, and applications will further advance biological research and clinical assessments.

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Section: Deconvolution Analysismentioning

confidence: 99%

Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications

et al. 2020

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“…We used a fast deconvolution algorithm based on a least-square curve-fitting method with a library of exponentials 33 . A model function with N lifetime components is defined as where b k (t) = exp(− t / τ k ); k = 1, 2 ⋯ N represents the library of exponential functions.…”

Section: Methodsmentioning

confidence: 99%

Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis

et al. 2021

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Total internal reflection fluorescence (TIRF) microscopy, which has about 100-nm axial excitation depth, is the method of choice for nanometer-sectioning imaging for decades. Lately, several new imaging techniques, such as variable angle TIRF microscopy, supercritical-angle fluorescence microscopy, and metal-induced energy transfer imaging, have been proposed to enhance the axial resolution of TIRF. However, all of these methods use high numerical aperture (NA) objectives, and measured images inevitably have small field-of-views (FOVs). Small-FOV can be a serious limitation when multiple cells need to be observed. We propose large-FOV nanometer-sectioning microscopy, which breaks the complementary relations between the depth of focus and axial sectioning by using MIET. Large-FOV imaging is achieved with a low-magnification objective, while nanometer-sectioning is realized utilizing metal-induced energy transfer and biexponential fluorescence lifetime analysis. The feasibility of our proposed method was demonstrated by imaging nanometer-scale distances between the basal membrane of human aortic endothelial cells and a substrate.

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“…The time the first photon arrives on the detector after excitation pulse is measured and stored in the memory so that the histogram of photon times and arrivals presents a curve of fluorescence intensity in terms of time [90][91][92] (Figure 3). Since the measurement data require time resolution of a few seconds, FlIM systems are equipped with high-speed supersensitive detectors capable of detecting single photons [93,94].…”

Section: Nadh and Fad Fluorescence Lifetime In Energy Metabolism Evalmentioning

confidence: 99%

Metabolic Imaging in the Study of Oncological Processes (Review)

Lukina¹,

Shirmanova²,

Sergeeva³

et al. 2016

Sovrem Tehnol Med

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There have been reviewed the main approaches to the study of energy metabolism in cancer cells, based on fluorescent imaging of NADH and FAD cofactors. The term "metabolic imaging" covers a range of modern fluorescent techniques for detecting NADH and FAD according to fluorescence intensity and/or lifetime. These cofactors play an important role in energy metabolism reactions acting as electron carriers and, being fluorescent, serve as a basis for metabolic process analysis in living cells and tissues without the use of additional coloring agents. Particular attention is paid to metabolic changes associated with carcinogenesis. Numerous examples of metabolic imaging application in cell cultures in vitro, animal and human tumors in vivo, as well as in patients' tumor biopsy samples have demonstrated its being highly demanded for biomedical research in the area of oncology.Key words: metabolic imaging; energy metabolism; fluorescence lifetime; redox ratio; NADH; FAD; oxidative phosphorylation; glycolysis; tumor cells.For contacts: Mariya M. lukina, e-mail: kuznetsova.m.m@yandex.ru Cells need energy to maintain homeostasis. All processes of cell activity, such as generating concentration gradients, cytoskeleton movement, DNA repair, transcription, translation and vesicular transport, are energy-dependent. Energy metabolism of normal cells is known to differ significantly from cell metabolism in case of a disease. Therefore, the metabolic status can serve as an indicator for diagnosis and visualization of response to pathological process treatment. Taking into account high incidence of oncological diseases, assessment of metabolic phenotype of tumor cells is particularly urgent.Development of optical visualization technologies has provided the possibility of noninvasive analysis of metabolic cofactors NADH and FAD in living tumor cells at high spatial resolution (up to several hundred Metabolic Imaging in the Study of Oncological Process nanometers) without using additional coloring agents and with no significant effect on biochemical and physiological condition of cells. The term "metabolic imaging" covers a range of modern fluorescent techniques which allow visualizing NADH and FAD according to their fluorescence intensity and/or lifetime.The present review characterizes the properties of energy metabolism in cancer cells, describes the two key approaches to the assessment of metabolic status: analysis of cofactor fluorescence intensity ratio (redox ratio) and their fluorescence lifetime. There have been presented numerous examples of metabolic imaging application in cell cultures in vitro, animal and human tumors in vivo, as well as in patients' tumor biopsy samples.

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Deconvolution of fluorescence lifetime imaging microscopy by a library of exponentials

Cited by 15 publications

References 26 publications

Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications

Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications

Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis

Metabolic Imaging in the Study of Oncological Processes (Review)

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