Fluorescence lifetime images and correlation spectra obtained by multidimensional time‐correlated single photon counting

Becker, Wolfgang; Bergmann, Axel; Haustein, Elke; Petrášek, Zdeněk; Schwille, Petra; Biskup, Christoph; Kelbauskas, Laimonas; Benndorf, Klaus; Klöcker, Nikolaj; Anhut, Tiemo; Riemann, Iris; König, Karsten

doi:10.1002/jemt.20251

Cited by 71 publications

(62 citation statements)

References 71 publications

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“…Considering the fact that many endogenous chromophores (quenched donors in FRET, NADH, NADPH) are characterised by fluorescence lifetimes in ps-range, the time-accuracy provided by FD FLIM is often insufficient. In contrast, TD FLIM techniques reach time-resolutions in the low ps-range or even fs-range [8][9][10][11]40,[45][46][47] .…”

Section: Time-domain Flimmentioning

confidence: 96%

“…FLIM in time-domain is primarily more advantageous than FLIM in frequency-domain because it provides a direct measurement of the fluorescence decay and, thus, a direct access to the parameter/parameters of interest 9,10,12,[40][41][42][43][44][45] . Considering the fact that many endogenous chromophores (quenched donors in FRET, NADH, NADPH) are characterised by fluorescence lifetimes in ps-range, the time-accuracy provided by FD FLIM is often insufficient.…”

Section: Time-domain Flimmentioning

confidence: 99%

“…It makes use of the fact that the detection of several photons in different detection channels in one laser period is unlikely. Therefore, the single photon pulses from all detector channels can be combined into a common photon pulse line and sent through the normal time measurement procedure of the TCSPC module 10,40,46 .…”

Section: Time-correlated Single Photon Counting (Tcspc)mentioning

confidence: 99%

“…The technique can reasonably be used for up to four individual ultrafast MCP-PMTs (e.g. R3809U by Hamamatsu) or one 16 channels or 32 channels multi-anode PMT 10,40,46 .…”

Section: Time-correlated Single Photon Counting (Tcspc)mentioning

confidence: 99%

“…homodyne and heterodyne frequency-domain FLIM 8 , time-correlated single-photon counting (TCSPC) 9,10 and time-gated techniques 11,12 as well as on their compatibility with current high-end far-field imaging techniques with particular relevance for the biosciences. The comparison criteria refer to standard fluorescence imaging parameters, i.e.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescence lifetime imaging in biosciences: technologies and applications

Niesner

Gericke

2008

Front. Phys. China

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The biosciences require the development of methods, which allow a non-invasive and rapid investigation of biological systems. In this frame high-end imaging techniques allow an intravital microscopy in real-time, providing information on a molecular basis. Far-field fluorescence imaging techniques are some of the most adequate methods for such investigations. However, there are great differences between the common fluorescence imaging techniques, i.e. wide-field, confocal one-photon and two-photon microscopy, respectively, as far as their applicability in diverse bioscientific research areas is concerned. In the first part of this work, we briefly compare these techniques. Standard methods used in the biosciences, i.e. steady-state techniques based on the analysis of the total fluorescence signal originating from the sample, can successfully be employed in the study of cell, tissue and organ morphology as well as in monitoring the macroscopic tissue function. However, they are mostly inadequate for the quantitative investigation of the cellular function at molecular level. The intrinsic disadvantages of steady-state techniques are counteracted by using time-resolved techniques. Among these the fluorescence lifetime imaging (FLIM) is currently the most common. Different FLIM principles as well as applications of particular relevance for the biosciences, especially for fast intravital studies are discussed in this work.

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Section: Time-domain Flimmentioning

confidence: 96%

Section: Time-domain Flimmentioning

confidence: 99%

Section: Time-correlated Single Photon Counting (Tcspc)mentioning

confidence: 99%

“…The technique can reasonably be used for up to four individual ultrafast MCP-PMTs (e.g. R3809U by Hamamatsu) or one 16 channels or 32 channels multi-anode PMT 10,40,46 .…”

Section: Time-correlated Single Photon Counting (Tcspc)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Fluorescence lifetime imaging in biosciences: technologies and applications

Niesner

Gericke

2008

Front. Phys. China

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Fluorescence

Birch¹,

Chen²,

Rolinski³

2015

Photonics

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This chapter surveys some of the main capabilities, techniques, and measurements that are enabled by fluorescence. It covers spectra, quantum yield, lifetime, quenching, anisotropy, and microscopy, in each case citing topical review articles, many of the original references, underlying theory and modern day applications. Measuring absorption and fluorescence spectra is usually the first place to start in any fluorescence study. In order to illustrate how absorption and fluorescence spectra often interplay in tandem, the chapter considers the example of the auto-oxidation of 3,4-dihydroxy-l-phenylalanine (l-DOPA) to produce melanin. Lifetime measurement has emerged in recent years as the most powerful and versatile technique in fluorescence spectroscopy. The basis of fluorescence anisotropy techniques is to use polarized excitation to create a spatially selected, non-random, distribution of excited fluorescent molecules which then randomize, most commonly by Brownian rotation, but also at times by energy migration depending on the system

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Fluorescence Imaging for Biomedical Analysis

Fixler

2015

The Optics Encyclopedia

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Fluorescence imaging techniques became powerful tools for inquisitive biologists and medical doctors. During the past decade, the description of new imaging modalities such as fluorescence life‐time, confocal, fluorescence anisotropy, and two‐photon microscopy has triggered an explosion in fluorescence imaging for biomedical analysis techniques. This article describes the basic principle of fluorescence imaging techniques and elaborates about their biomedical analysis applications

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Fluorescence lifetime images and correlation spectra obtained by multidimensional time‐correlated single photon counting

Cited by 71 publications

References 71 publications

Fluorescence lifetime imaging in biosciences: technologies and applications

Fluorescence lifetime imaging in biosciences: technologies and applications

Fluorescence

Fluorescence Imaging for Biomedical Analysis

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