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Effect of probe diffusion on the SOFI imaging accuracy.

ABSTRACT: Live-cell super-resolution fluorescence imaging is becoming commonplace for exploring biological systems, though sample dynamics can affect the imaging quality. In this work we evaluate the effect of probe diffusion on super-resolution optical fluctuation imaging (SOFI), using a theoretical model and numerical simulations based on the imaging of live cells labelled with photochromic fluorescent proteins. We find that, over a range of physiological conditions, fluorophore diffusion results in a change in the amplitude of the SOFI signal. The magnitude of this change is approximately proportional to the on-time ratio of the fluorophores. However, for photochromic fluorescent proteins this effect is unlikely to present a significant distortion in practical experiments in biological systems. Due to this lack of distortions, probe diffusion strongly enhances the SOFI imaging by avoiding spatial undersampling caused by the limited labeling density.

SUBMITTER: Vandenberg W

PROVIDER: S-EPMC5363082 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Effect of probe diffusion on the SOFI imaging accuracy.

Vandenberg Wim W Dedecker Peter P

Scientific reports 20170323

Live-cell super-resolution fluorescence imaging is becoming commonplace for exploring biological systems, though sample dynamics can affect the imaging quality. In this work we evaluate the effect of probe diffusion on super-resolution optical fluctuation imaging (SOFI), using a theoretical model and numerical simulations based on the imaging of live cells labelled with photochromic fluorescent proteins. We find that, over a range of physiological conditions, fluorophore diffusion results in a c ...[more]

PMID: 28333166

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Project description:To evaluate the incremental value of diffusion-weighted (DW) imaging and apparent diffusion coefficient (ADC) mapping in relation to conventional breast magnetic resonance (MR) imaging in the characterization of benign versus malignant breast lesions at 3.0 T.This retrospective HIPAA-compliant study was approved by the institutional review board, with the requirement for informed patient consent waived. Of 550 consecutive patients who underwent bilateral breast MR imaging over a 10-month period, 93 women with 101 lesions met the following study inclusion criteria: They had undergone three-dimensional (3D) high-spatial-resolution T1-weighted contrast material-enhanced MR imaging, dynamic contrast-enhanced MR imaging, and DW imaging examinations at 3.0 T and either had received a pathologic analysis-proven diagnosis (96 lesions) or had lesion stability confirmed at more than 2 years of follow-up (five lesions). DW images were acquired with b values of 0 and 600 sec/mm(2). Regions of interest were drawn on ADC maps of breast lesions and normal glandular tissue. Morphologic features (margin, enhancement pattern), dynamic contrast-enhanced MR results (semiquantitative kinetic curve data), absolute ADCs, and glandular tissue-normalized ADCs were included in multivariate models to predict a diagnosis of benign versus malignant lesion.Forty-one (44%) of the 93 patients were premenopausal, and 52 (56%) were postmenopausal. Thirty-three (32.7%) of the 101 lesions were benign, and 68 (67.3%) were malignant. Normalized ADCs were significantly different between the benign (mean ADC, 1.1 x 10(-3) mm(2)/sec +/- 0.4 [standard deviation]) and malignant (mean ADC, 0.55 x 10(-3) mm(2)/sec +/- 0.16) lesions (P < .001). Adding normalized ADCs to the 3D T1-weighted and dynamic contrast-enhanced MR data improved the diagnostic performance of MR imaging: The area under the receiver operating characteristic curve improved from 0.89 to 0.98, and the false-positive rate decreased from 36% (nine of 25 lesions) to 24% (six of 25 lesions).DW imaging with glandular tissue-normalized ADC assessment improves the characterization of breast lesions beyond the characterization achieved with conventional 3D T1-weighted and dynamic contrast-enhanced MR imaging at 3.0 T.

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Effect of probe diffusion on the SOFI imaging accuracy.

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Effect of probe diffusion on the SOFI imaging accuracy.

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