Fluorescence Lifetime Imaging (FLIM): Basic Concepts and Recent Applications

Suhling, Klaus; Hirvonen, L; Levitt, James A.; Chung, Pei‐Hua; Tregido, Carolyn; Marois, Alix Le; Rusakov, Dmitri A.; Zheng, Kaiyu; Ameer-Beg, Simon; Poland, Simon P.; Coelho, Simao; Dimble, Richard

doi:10.1007/978-3-319-14929-5_3

Cited by 18 publications

(14 citation statements)

References 450 publications

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“…Pump-probe microscopy is an adaptation of transient absorption spectroscopy [8,9] that uses low pulse-energy, high repetition-rate ultrafast laser sources and a few selected pump/probe wavelength combinations at a time, trading broad spectral coverage for fast, high resolution, 3-dimensional imaging capabilities. Pump-probe is somewhat akin to fluorescence lifetime microscopy [10,11], but without the need for appreciable fluorescence quantum yield. Rather, pump-probe resolves excited-state lifetimes by measuring subtle changes in optical absorption of a probe pulse following absorption of a pump pulse.…”

Section: Introductionmentioning

confidence: 99%

Transient absorption imaging of hemes with 2-color, independently tunable visible-wavelength ultrafast source

Domingue¹,

Bartels²,

Chicco³

et al. 2017

Biomed. Opt. Express

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Pump probe microscopy is a time-resolved multiphoton imaging technique capable of generating contrast between non-fluorescent pigments based on differences in excited-state lifetimes. Here we describe a fiber-based ultrafast system designed for imaging heme proteins with an independently-tunable pulse pair in the visible-wavelength regime. Starting with a 1060 nm fiber amplifier (1.3 W at 63 MHz, 140 fs pulses), visible pulses were produced in the vicinity of 488 nm and 532 nm by doubling the output of a short photonic crystal fiber with a pair of periodically-poled lithium niobate crystals, providing 5-20 mW power in each beam. This was sufficient for acquiring transient absorption images from unstained cryosectioned tissue.

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Section: Introductionmentioning

confidence: 99%

Transient absorption imaging of hemes with 2-color, independently tunable visible-wavelength ultrafast source

Domingue¹,

Bartels²,

Chicco³

et al. 2017

Biomed. Opt. Express

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“…TCSPC-based FLIM has the best signal-to-noise ratio of any FLIM techniques and it is an accepted gold standard for FLT measurement. Here we show the schematic of a typical TCSPC-based FLIM system 31,32 in Fig. 5.…”

Section: Fluorescence Lifetime Imaging Microscopy Systemmentioning

confidence: 99%

Fluorescence lifetime imaging microscopy and its applications in skin cancer diagnosis

Liu

Yang

Zhang

et al. 2019

J. Innov. Opt. Health Sci.

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Fluorescence lifetime (FLT) of fluorophores is sensitive to the changes in their surrounding microenvironment, and hence it can quantitatively reveal the physiological characterization of the tissue under investigation. Fluorescence lifetime imaging microscopy (FLIM) provides not only morphological but also functional information of the tissue by producing spatially resolved image of fluorophore lifetime, which can be used as a signature of disorder and/or malignancy in diseased tissues. In this paper, we begin by introducing the basic principle and common detection methods of FLIM. Then the recent advances in the FLIM-based diagnosis of three different skin cancers, including basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and malignant melanoma (MM) are reviewed. Furthermore, the potential advantages of FLIM in skin cancer diagnosis and the challenges that may be faced in the future are prospected.

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“…[3][4][5][6][7][8][9] It can thus be conveniently used to achieve quantitative measurements and to distinguish di®erent probes with similar°uorescence spectra. Importantly, according to the relationship between°uorescence lifetime and the environment around°uorescent molecules, FLIM technology can favorably be used to quantitatively measure various biochemical parameters in the microenvironment, 6 such as oxygen content, 5,10,11 pH, 12,13 metabolic state, 10,14,15 viscosity, 16,17 solution hydrophobicity, ion concentrations, 18,19 quencher distribution, temperature, 20 and°uorescence resonance energy transfer (FRET) e±ciency. [21][22][23][24] Therefore, FLIM has played an increasingly important role in the biomedical¯eld in the past two decades.…”

Section: Introductionmentioning

confidence: 99%

Fast fluorescence lifetime imaging techniques: A review on challenge and development

Liu

Lin

Becker

et al. 2019

J. Innov. Opt. Health Sci.

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Fluorescence lifetime imaging microscopy (FLIM) is increasingly used in biomedicine, material science, chemistry, and other related research fields, because of its advantages of high specificity and sensitivity in monitoring cellular microenvironments, studying interaction between proteins, metabolic state, screening drugs and analyzing their efficacy, characterizing novel materials, and diagnosing early cancers. Understandably, there is a large interest in obtaining FLIM data within an acquisition time as short as possible. Consequently, there is currently a technology that advances towards faster and faster FLIM recording. However, the maximum speed of a recording technique is only part of the problem. The acquisition time of a FLIM image is a complex function of many factors. These include the photon rate that can be obtained from the sample, the amount of information a technique extracts from the decay functions, the efficiency at which it determines fluorescence decay parameters from the recorded photons, the demands for the accuracy of these parameters, the number of pixels, and the lateral and axial resolutions that are obtained in biological materials. Starting from a discussion of the parameters which determine the acquisition time, this review will describe existing and emerging FLIM techniques and data analysis algorithms, and analyze their performance and recording speed in biological and biomedical applications.

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Fluorescence Lifetime Imaging (FLIM): Basic Concepts and Recent Applications

Cited by 18 publications

References 450 publications

Transient absorption imaging of hemes with 2-color, independently tunable visible-wavelength ultrafast source

Transient absorption imaging of hemes with 2-color, independently tunable visible-wavelength ultrafast source

Fluorescence lifetime imaging microscopy and its applications in skin cancer diagnosis

Fast fluorescence lifetime imaging techniques: A review on challenge and development

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