Widefield multifrequency fluorescence lifetime imaging using a two‐tap complementary metal‐oxide semiconductor camera with lateral electric field charge modulators

Chen, Hongtao; Ma, Ning; Kagawa, Keiichiro; Kawahito, Shoji; Digman, Michelle A.; Gratton, Enrico

doi:10.1002/jbio.201800223

Cited by 6 publications

(3 citation statements)

References 34 publications

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“…136,[140][141][142][143] Other advantages of wide-field systems are their simpler implementation and the low computational cost to assign photon detection times in each pixel. Some of the advancements in wide-field FLIM include its implementation with Nipkow disc microscopy for fast 3-D FLIM imaging, 142 wide-field coupled with single plane illumination microscopy for high-resolution 3-D FLIM, 144 TG single photon avalanche diode (SPAD) cameras for phasor-based high speed wide-field FLIM, 145 multifrequency widefield, 146,147 and image gating by pockel cells. 148 Current wide-field FLIM systems are discussed in detail by Suhling and Hirvonen et al 149…”

Section: Microscopymentioning

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: Microscopymentioning

confidence: 99%

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

et al. 2020

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“…The acquisition speed of FD-FLIM is comparable to fast time-domain implementations [79,80]. FD-FLIM at ∼24 Hz using time-of-flight detection was demonstrated in 2004 [74].…”

Section: Frequency Domain Flimmentioning

confidence: 99%

High-throughput, multi-parametric, and correlative fluorescence lifetime imaging

Poudel

Mela

Kaminski

2020

Methods Appl. Fluoresc.

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In this review, we discuss methods and advancements in fluorescence lifetime imaging microscopy that permit measurements to be performed at faster speed and higher resolution than previously possible. We review fast single-photon timing technologies and the use of parallelized detection schemes to enable high-throughput and high content imaging applications. We appraise different technological implementations of fluorescence lifetime imaging, primarily in the time-domain. We also review combinations of fluorescence lifetime with other imaging modalities to capture multi-dimensional and correlative information from a single sample. Throughout the review, we focus on applications in biomedical research. We conclude with a critical outlook on current challenges and future opportunities in this rapidly developing field.

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“…Measurements of fluorescence lifetime are not affected by the fluorescence probe concentration, excitation light intensity, photobleaching, or other factors, so they have the advantages of strong specificity, high sensitivity, and the ability to perform quantitative measurements [ 41 , 42 ]. In recent years, FLIM technology has enabled important research progress in practical applications for biomedicine, material science, and many other fields [ 43 , 44 , 45 , 46 ]. FLIM technology is often used to detect the distribution of cell microenvironmental parameters and the state of energy metabolism, as well as to characterize the internal structure and properties of materials [ 47 ].…”

Section: Introductionmentioning

confidence: 99%

A Rapid Method for Detecting Microplastics Based on Fluorescence Lifetime Imaging Technology (FLIM)

Fang

Wang

et al. 2022

Toxics

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With the increasing use and release of plastic products, microplastics have rapidly accumulated in ecological environments. When microplastics enter the food chain, they cause serious harm to organisms and humans. Microplastics pollution has become a growing concern worldwide; however, there is still no standardized method for rapidly and accurately detecting microplastics. In this work, we used fluorescence lifetime imaging technology to detect four kinds of Nile red-stained and unstained microplastics, and the unique phasor fingerprints of different microplastics were obtained by phasor analysis. Tracing the corresponding pixels of the “fingerprint” in the fluorescence lifetime image allowed for the quick and intuitive identification of different microplastics and their location distributions in a mixed sample. In our work, compared with staining the four microplastics with a fluorescent dye, using the phasor “fingerprint library” formed by the autofluorescence lifetimes of the microplastics was more easily distinguished than microplastics in the mixed samples. The feasibility of this method was further tested by adding three single substances—SiO2, chitin and decabromodiphenyl ethane (DBDPE), and surface sediments to simulate interferent in the environment, and the results providing potential applications for the identification and analysis of microplastics in complex environments.

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Widefield multifrequency fluorescence lifetime imaging using a two‐tap complementary metal‐oxide semiconductor camera with lateral electric field charge modulators

Cited by 6 publications

References 34 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

High-throughput, multi-parametric, and correlative fluorescence lifetime imaging

A Rapid Method for Detecting Microplastics Based on Fluorescence Lifetime Imaging Technology (FLIM)

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