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STED microscope with spiral phase contrast.

ABSTRACT: Stimulated Emission Depletion (STED) microscopy enables superresolution imaging of fluorescently marked nano-structures in vivo. Biological investigations are often hindered by the difficulty of relating super-resolved structures to other non-labeled features. Here we demonstrate that the similarity in optical design of Spiral Phase Contrast (SPC) and STED microscopes allows straightforward implementation of a phase contrast channel into a STED microscope in widefield and scanning modes. This method allows dual imaging and overlay in two contrast modes in fixed and in living specimens, in which double labeling is especially challenging. Living GFP- and YPF-stained neurons are imaged in one label-free phase contrast and one high-resolution STED channel. Furthermore, we implement SPC in widefield and scanning modes demonstrating that scanning confocal SPC yields the highest optical contrast. The latter configuration can provide contour detection or highlights and shadows reminiscent of differential interference contrast.

SUBMITTER: Lauterbach MA

PROVIDER: S-EPMC3689173 | biostudies-other | 2013

REPOSITORIES: biostudies-other

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STED microscope with spiral phase contrast.

Lauterbach Marcel A MA Guillon Marc M Soltani Asma A Emiliani Valentina V

Scientific reports 20130101

Stimulated Emission Depletion (STED) microscopy enables superresolution imaging of fluorescently marked nano-structures in vivo. Biological investigations are often hindered by the difficulty of relating super-resolved structures to other non-labeled features. Here we demonstrate that the similarity in optical design of Spiral Phase Contrast (SPC) and STED microscopes allows straightforward implementation of a phase contrast channel into a STED microscope in widefield and scanning modes. This me ...[more]

PMID: 23787399

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Project description:To validate a prospectively triggered spiral phase-contrast magnetic resonance (MR) sequence accelerated with sensitivity encoding (SENSE) in a population of children and adults with congenital heart disease.The local research ethics committee approved this study, and written consent was obtained from all patients or guardians. Stroke volumes were quantified in 40 patients (mean age ± standard deviation: 21.4 years ± 13.8, age range: 3.0-61.3 years; 22 male patients aged 3.0-38.0 years [mean age, 17.2 years ± 10.5], 18 female patients aged 4.7-61.3 years [mean age, 26.6 years ± 15.9]) with congenital heart disease in the aorta (n = 40), main pulmonary artery (n = 38), right pulmonary artery (n = 22), and left pulmonary artery (n = 24). Stroke volumes were obtained with (a) breath-hold spiral phase-contrast MR imaging with SENSE, (b) conventional breath-hold cartesian phase-contrast MR imaging, and (c) reference free-breathing phase-contrast MR imaging. Stroke volumes were compared by using repeated-measures analysis of variance, Bland-Altman analysis, and correlation coefficients.Imaging time with the breath-hold spiral phase-contrast MR sequence was significantly lower than that with the conventional breath-hold phase-contrast MR sequence (~5 seconds vs ~16 seconds, respectively; P < .0001). There was excellent agreement in stroke volumes in all vessels between the reference free-breathing sequence (mean volume, 60.3 mL ± 27.3) and the two breath-hold sequences-spiral SENSE phase-contrast MR imaging (mean volume, 59.5 mL ± 27.1; P < .001) and conventional cartesian phase-contrast MR imaging (mean volume, 59.8 mL ± 27.6; P = .268). The limits of agreement were smaller with the spiral breath-hold sequence than with the conventional breath-hold sequence (-4.4 mL, 2.9 mL vs -10.3 mL, 9.3 mL, respectively); correlation was similar (r = 0.998 vs r = 0.984, respectively).Flow volumes can be accurately and reliably quantified by using a spiral SENSE phase-contrast MR sequence, with high spatiotemporal resolution obtained in a short breath hold.

Dataset Information

STED microscope with spiral phase contrast.

Publications

STED microscope with spiral phase contrast.

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