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MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution.

ABSTRACT: Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ?2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution.

SUBMITTER: Eilers Y

PROVIDER: S-EPMC6004438 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution.

Eilers Yvan Y Ta Haisen H Gwosch Klaus C KC Balzarotti Francisco F Hell Stefan W SW

Proceedings of the National Academy of Sciences of the United States of America 20180529 24

Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 milli ...[more]

PMID: 29844182

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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.

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MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution.

Publications

MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution.

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