Real-time time-domain fluorescence lifetime imaging including single-shot acquisition with a segmented optical image intensifier

Elson, Daniel S.; Munro, Ian; Requejo-Isidro, José; McGinty, James; Dunsby, C; Galletly, Neil; Stamp, Gordon W.; Neil, Mark A. A.; Lever, M. J.; Kellett, P.A.; Dymoke‐Bradshaw, A. K. L.; Hares, J. D.; French, Paul M. W.

doi:10.1088/1367-2630/6/1/180

Cited by 74 publications

(62 citation statements)

References 24 publications

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“…Another way to increase fluorescence signals is to use lower excitation intensity coupled with longer image acquisition times, but sample movement will be a major concern of this approach in livecell imaging. 8 Considering the above challenges, a combination of low fluorescence signals, low excitation light intensity, and fast image acquisition can make FLIM data very imprecise in biological applications.…”

Section: Introductionmentioning

confidence: 99%

“…To improve the precision of lifetime determination in FLIM, error analysis with Monte Carlo ͑MC͒ simulations may be used to determine optimal gating schemes. [8][9][10][11][12] Optimal gating schemes of rapid lifetime determination ͑RLD͒ for single-exponential decays ͑a two-gate protocol͒ with respect to relative standard deviation ͓͑RSD͒, also commonly known as coefficient of variation͔ of lifetime have been reported, 10,12 and an error analysis of RLD for double-exponential decays has also been addressed. 11 Recently, optimization of fluorescence lifetime sensing in frequency domain was studied.…”

Section: Introductionmentioning

confidence: 99%

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Enhancing precision in time-domain fluorescence lifetime imaging

Chang

Mycek

2010

J. Biomed. Opt.

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Abstract. In biological applications of fluorescence lifetime imaging, low signals from samples can be a challenge, causing poor lifetime precision. We demonstrate how optimal signal gating ͑a method applied to the temporal dimension of a lifetime image͒ and novel total variation denoising models ͑a method applied to the spatial dimension of a lifetime image͒ can be used in time-domain fluorescence lifetime imaging microscopy ͑FLIM͒ to improve lifetime precision. In time-gated FLIM, notable fourfold precision improvements were observed in a low-light example. This approach can be employed to improve FLIM data while minimizing sample light exposure and increasing imaging speed.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Enhancing precision in time-domain fluorescence lifetime imaging

Chang

Mycek

2010

J. Biomed. Opt.

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“…Nevertheless, using an optical delay line will decrease the signal intensity since the fluorescence signal is not collimated. Another way to obtain single-shot lifetime images would be to use a multi-channel segmented gated optical intensifier, such as the one used by Elson et al [19] for sequential time gated FLI. By using DIME, the signal-to-noise ratio could be improved significantly as compared to using sequential time gating.…”

Section: Discussionmentioning

confidence: 99%

“…However, this time measured lifetimes were longer than 30 ns [10]. Microscopic FLI investigations in the field of biomedicine have been performed in the time domain by Elson et al [19]. In this work fluorescence lifetimes of a few nanoseconds were measured in single acquisition with a repetition rate of 20 fps, corresponding to an acquisition time of 50 ms.…”

Section: Introductionmentioning

confidence: 97%

Single-laser shot fluorescence lifetime imaging on the nanosecond timescale using a Dual Image and Modeling Evaluation algorithm

Ehn¹,

Johansson²,

Arvidsson³

et al. 2012

Opt. Express

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“…An on-board field-programmable gate array or a computer can process this information to provide distance measurements. ToF technologies can then provide single-shot real-time imaging systems for high speed FLIM, an application that is currently only available in specialist laboratories [8,33,34]. …”

Section: Beyond Rangementioning

confidence: 99%

Beyond Range: Innovating Fluorescence Microscopy

Esposito

2012

Remote Sensing

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Time-of-Flight (ToF) technologies are developed mainly for range estimations in industrial applications or consumer products. Recently, it was realized that ToF sensors could also be used for the detection of fluorescence and of the minute changes in the nanosecond-lived electronic states of fluorescent molecules. This capability can be exploited to report on the biochemical processes occurring within living organisms. ToF technologies, therefore, provide new opportunities in molecular and cell biology, diagnostics, and drug discovery. In this short communication, the convergence of the engineering and biomedical communities onto ToF technologies and its potential impact on basic, applied and translational sciences are discussed.Keywords: Time-of-Flight (ToF); fluorescence lifetime; microscopy Converging CommunitiesIt was the summer of 2005 when in the laboratory of Fred Wouters we started the first autonomous runs of our new in-house developed high throughput fluorescence lifetime imaging (FLIM) system. However successful that development was, one key element in allowing the biomedical community to benefit from FLIM systems, to enable the screening technologies we were pioneering [1] and to get closer to the routine application of FLIM, was still missing: replacing the expensive, comparatively fragile, and not very user-friendly, multi-channel plates used in FLIM with better technologies. The basic idea was to find a silicon-based detector capable of demodulating fluorescence signals on a bi-dimensional array of pixels. Frequent searches of primary literature returned only a few, though interesting, papers including: Nishikata et al. [2] on the development of a dedicated lock-in imager in CCD technology and, Mitchell et al. [3,4] on the adaptation of conventional CCD cameras to achieve demodulation. Unfortunately, ad hoc technologies for lock-in imaging appeared to be operating at OPEN ACCESS

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Real-time time-domain fluorescence lifetime imaging including single-shot acquisition with a segmented optical image intensifier

Cited by 74 publications

References 24 publications

Enhancing precision in time-domain fluorescence lifetime imaging

Enhancing precision in time-domain fluorescence lifetime imaging

Single-laser shot fluorescence lifetime imaging on the nanosecond timescale using a Dual Image and Modeling Evaluation algorithm

Beyond Range: Innovating Fluorescence Microscopy

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