Dataset Information

Ultra-High Resolution 3D Imaging of Whole Cells.

ABSTRACT: Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. We demonstrate the wide applicability of W-4PiSMSN across diverse research fields by imaging complex molecular architectures ranging from bacteriophages to nuclear pores, cilia, and synaptonemal complexes in large 3D cellular volumes.

SUBMITTER: Huang F

PROVIDER: S-EPMC5005454 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature

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Publications

Ultra-High Resolution 3D Imaging of Whole Cells.

Huang Fang F Sirinakis George G Allgeyer Edward S ES Schroeder Lena K LK Duim Whitney C WC Kromann Emil B EB Phan Thomy T Rivera-Molina Felix E FE Myers Jordan R JR Irnov Irnov I Lessard Mark M Zhang Yongdeng Y Handel Mary Ann MA Jacobs-Wagner Christine C Lusk C Patrick CP Rothman James E JE Toomre Derek D Booth Martin J MJ Bewersdorf Joerg J

Cell 20160707 4

Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of ...[more]

PMID: 27397506

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Project description:BackgroundConventional 2D inversion recovery (IR) and phase sensitive inversion recovery (PSIR) late gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR) have been widely incorporated into routine CMR for the assessment of myocardial viability. However, reliable suppression of fat signal, and increased isotropic spatial resolution and volumetric coverage within a clinically feasible scan time remain a challenge. In order to address these challenges, this work proposes a highly efficient respiratory motion-corrected 3D whole-heart water/fat LGE imaging framework.MethodsAn accelerated IR-prepared 3D dual-echo acquisition and motion-corrected reconstruction framework for whole-heart water/fat LGE imaging was developed. The acquisition sequence includes 2D image navigators (iNAV), which are used to track the respiratory motion of the heart and enable 100% scan efficiency. Non-rigid motion information estimated from the 2D iNAVs and from the data itself is integrated into a high-dimensional patch-based undersampled reconstruction technique (HD-PROST), to produce high-resolution water/fat 3D LGE images. A cohort of 20 patients with known or suspected cardiovascular disease was scanned with the proposed 3D water/fat LGE approach. 3D water LGE images were compared to conventional breath-held 2D LGE images (2-chamber, 4-chamber and stack of short-axis views) in terms of image quality (1: full diagnostic to 4: non-diagnostic) and presence of LGE findings.ResultsImage quality was considered diagnostic in 18/20 datasets for both 2D and 3D LGE magnitude images, with comparable image quality scores (2D: 2.05?±?0.72, 3D: 1.88?±?0.90, p-value?=?0.62) and overall agreement in LGE findings. Acquisition time for isotropic high-resolution (1.3mm3) water/fat LGE images was 8.0?±?1.4?min (3-fold acceleration, 60-88 slices covering the whole heart), while 2D LGE images were acquired in 5.6?±?2.2?min (12-18 slices, including pauses between breath-holds) albeit with a lower spatial resolution (1.40-1.75?mm in-plane ×?8?mm slice thickness).ConclusionA novel framework for motion-corrected whole-heart 3D water/fat LGE imaging has been introduced. The method was validated in patients with known or suspected cardiovascular disease, showing good agreement with conventional breath-held 2D LGE imaging, but offering higher spatial resolution, improved volumetric coverage and good image quality from a free-breathing acquisition with 100% scan efficiency and predictable scan time.

Dataset Information

Ultra-High Resolution 3D Imaging of Whole Cells.

Publications

Ultra-High Resolution 3D Imaging of Whole Cells.

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