Adaptive wide-field optical tomography

Venugopal, Vivek; Intes, Xavier

doi:10.1117/1.jbo.18.3.036006

Cited by 36 publications

(30 citation statements)

References 32 publications

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“…In this work, we used a wide-field time-gated imaging platform working in the NIR range 30 and employed an active illumination module. [31][32][33] This system has been designed to quantitatively image FRET donor fractions in vitro, ex vivo, and in vivo. 29,31,34 Briefly, the system uses a Maitai laser as a source and is coupled to a digital micromirror unit (DLP Discovery 4100 Kit and D2D module, Texas Instruments Inc., Dallas, Texas) to obtain a spatially controllable wide-field excitation [or active wide-field illumination (AWFI)].…”

Section: Time-domain Wide-field Gated Imagingmentioning

confidence: 99%

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer

et al. 2014

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Abstract. Fluorescence lifetime imaging (FLIM) aims at quantifying the exponential decay rate of fluorophores to yield lifetime maps over the imaged sample. When combined with Förster resonance energy transfer (FRET), the technique can be used to indirectly sense interactions at the nanoscale such as protein-protein interactions, protein-DNA interactions, and protein conformational changes. In the case of FLIM-FRET, the fluorescence intensity decays are fitted to a biexponential model in order to estimate the lifetime and fractional amplitude coefficients of each component of the population of the donor fluorophore (quenched and nonquenched). Numerous time data points, also called temporal or time gates, are typically employed for accurately estimating the model parameters, leading to lengthy acquisition times and significant computational demands. This work investigates the effect of the number and location of time gates on model parameter estimation accuracy. A detailed model of a FLIM-FRET imaging system is used for the investigation, and the simulation outcomes are validated with in vitro and in vivo experimental data. In all cases investigated, it is found that 10 equally spaced time gates allow robust estimation of model-based parameters with accuracy similar to that of full temporal datasets (90 gates).

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Section: Time-domain Wide-field Gated Imagingmentioning

confidence: 99%

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer

et al. 2014

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“…Fig. 2(b)) whereas they were within 10% of accuracy when using the pico-projector [14]. Moreover, the overall spatial uniformity of the imaging platform, as measured as by imaging a homogenous input pattern, was characterized at ~85% [11].…”

Section: Calibration Of Active Illumination Modulementioning

confidence: 84%

“…However, it is worth noting that this approach can be readily translated to in vivo fluorescence molecular tomography (FMT). In our previous work [14], we have implemented AWFI to the excitation field of a synthetic animal model for FMT to increase SNR and the tomographic information. The AWFI described in this paper has additional advantages.…”

Section: Active Illumination For In Vivo Biodistributionmentioning

confidence: 99%

Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging

Zhao¹,

Abe²,

Rajoria³

et al. 2014

Biomed. Opt. Express

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Fluorescence lifetime imaging is playing an increasing role in drug development by providing a sensitive method to monitor drug delivery and receptor-ligand interactions. However, the wide dynamic range of fluorescence intensity emitted by ex vivo and in vivo samples presents challenges in retrieving information over the whole subject accurately and quantitatively. To overcome this challenge, we developed an active widefield illumination (AWFI) strategy based on a spatial light modulator that acquires optimal fluorescence signals by enhancing the dynamic range, signal to noise ratio, and estimation of lifetime-based parameters. We demonstrate the ability of AWFI to estimate Förster resonance energy transfer (FRET) donor fraction from dissected organs with high accuracy (standard deviation <6%) over the whole field of view, in contrast with the homogenous wide-field illumination. We further report its successful application to quantitative FRET imaging in a live mouse. AWFI allows improved detection of weak signals and enhanced quantitative accuracy in ex vivo and in vivo molecular fluorescence quantitative imaging. The technique allows for robust quantitative estimation of the bio-distribution of molecular probes and lifetime-based parameters over an extended imaging field exhibiting a large range of fluorescence intensities and at a high acquisition speed (less than 1 min).

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“…While structured illumination FT systems can offer advantages over single-point illumination FT, they require more complex hardware that can pose challenges and complications for arbitrary non-flat subject geometries [30][31][32][33][34][35][36]. The advantage of Hadamard multiplexed FT is that it offers high robustness and high-throughput wide-field illumination similar to structured-illumination FT without the complex hardware requirements.…”

Section: Discussionmentioning

confidence: 99%

Hadamard multiplexed fluorescence tomography

Behrooz

Eftekhar

Adibi

2014

Biomed. Opt. Express

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Depth-resolved three-dimensional (3D) reconstruction of fluorophore-tagged inclusions in fluorescence tomography (FT) poses a highly ill-conditioned problem as depth information must be extracted from boundary data. Due to the ill-posed nature of the FT inverse problem, noise and errors in the data can severely impair the accuracy of the 3D reconstructions. The signal-to-noise ratio (SNR) of the FT data strongly affects the quality of the reconstructions. Additionally, in FT scenarios where the fluorescent signal is weak, data acquisition requires lengthy integration times that result in excessive FT scan periods. Enhancing the SNR of FT data contributes to the robustness of the 3D reconstructions as well as the speed of FT scans. A major deciding factor in the SNR of the FT data is the power of the radiation illuminating the subject to excite the administered fluorescent reagents. In existing single-point illumination FT systems, the source power level is limited by the skin maximum radiation exposure levels. In this paper, we introduce and study the performance of a multiplexed fluorescence tomography system with orders-of-magnitude enhanced data SNR over existing systems. The proposed system allows for multi-point illumination of the subject without jeopardizing the information content of the FT measurements and results in highly robust reconstructions of fluorescent inclusions from noisy FT data. Improvements offered by the proposed system are validated by numerical and experimental studies. tomography of sub-surface heterogeneities using spatially modulated structured light," Opt.

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Adaptive wide-field optical tomography

Cited by 36 publications

References 32 publications

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer

Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging

Hadamard multiplexed fluorescence tomography

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