Nanoscopy of bacterial cells immobilized by holographic optical tweezers

Diekmann, Robin; Wolfson, Deanna L.; Spahn, Christoph; Heilemann, Mike; Schüttpelz, Mark; Huser, Thomas

doi:10.1038/ncomms13711

Cited by 69 publications

(35 citation statements)

References 34 publications

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“…1), allowing the DNA to be elongated to ~95% of its theoretical length. Protein molecules were fluorescently labeled with ATTO 647N, and the emission signal was detected using a high light collection efficiency optical setup 38 combined with effective noise reduction strategies, including surface passivation and illumination of a thin layer of the sample 39 . Once DNA was localized and linearized (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V

et al. 2018

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In order to preserve genomic stability, cells rely on various repair pathways for removing DNA damage. The mechanisms how enzymes scan DNA and recognize their target sites are incompletely understood. Here, by using high-localization precision microscopy along with 133 Hz high sampling rate, we have recorded EndoV and OGG1 interacting with 12-kbp elongated λ-DNA in an optical trap. EndoV switches between three distinct scanning modes, each with a clear range of activation energy barriers. These results concur with average diffusion rate and occupancy of states determined by a hidden Markov model, allowing us to infer that EndoV confinement occurs when the intercalating wedge motif is involved in rigorous probing of the DNA, while highly mobile EndoV may disengage from a strictly 1D helical diffusion mode and hop along the DNA. This makes EndoV the first example of a monomeric, single-conformation and single-binding-site protein demonstrating the ability to switch between three scanning modes.

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Section: Resultsmentioning

confidence: 99%

“…Individual flow chambers were fixed onto the piezo-steerable stage of a custom-built combined holographic optical tweezer and subdiffraction resolution microscope which has previously been described 38 . In brief, a spatial light modulator was used to steer the infrared trapping laser within the sample.…”

Section: Methodsmentioning

confidence: 99%

Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V

et al. 2018

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“…By using a pre-designed optical set-up, left-handed or right-handed DNA/RNA-like molecules can be sorted if they are rotated in different directions, making it is possible to sort and manipulate individual DNA molecules ( Figure 8a ). Holographic optical tweezers can control the orientation of a cell at will and can achieve super-resolution using localization microscopy, yielding multiple perspectives of one sample with nanoscale localization precision( Figure 8b ) 194 . The new developments will lead to another revolution in OM and create abundant opportunities in bioscience 195 .…”

Section: Applications In Biochemical Manipulationmentioning

confidence: 99%

Optical manipulation from the microscale to the nanoscale: fundamentals, advances and prospects

et al. 2017

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Since the invention of optical tweezers, optical manipulation has advanced significantly in scientific areas such as atomic physics, optics and biological science. Especially in the past decade, numerous optical beams and nanoscale devices have been proposed to mechanically act on nanoparticles in increasingly precise, stable and flexible ways. Both the linear and angular momenta of light can be exploited to produce optical tractor beams, tweezers and optical torque from the microscale to the nanoscale. Research on optical forces helps to reveal the nature of light–matter interactions and to resolve the fundamental aspects, which require an appropriate description of momenta and the forces on objects in matter. In this review, starting from basic theories and computational approaches, we highlight the latest optical trapping configurations and their applications in bioscience, as well as recent advances down to the nanoscale. Finally, we discuss the future prospects of nanomanipulation, which has considerable potential applications in a variety of scientific fields and everyday life.

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“…However, even though these techniques are capable of high lateral resolution , they come with low axial resolution due to the missing angular coverage which can only partially be addressed by regularization. To record a sinogram with an angular coverage of 180° and above, multiple techniques, including optical tweezers , flow‐induced rotation in a microfluidic channel , and the mechanical rotation of the specimen or the imaging device , were proposed. In the present study, we combine a 360° contact‐free, optofluidic rotation of single, suspended cells with quantitative phase and fluorescence imaging.…”

Section: Introductionmentioning

confidence: 99%

Three‐dimensional correlative single‐cell imaging utilizing fluorescence and refractive index tomography

Schürmann

Cojoc

Girardo

et al. 2017

Journal of Biophotonics

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This is the pre-peer reviewed version of the following article: 3D correlative single-cell imaging utilizing fluorescence and refractive index tomography, which has been published in final form at https://dx.doi.org/ 10.1002/jbio.201700145.Cells alter the path of light, a fact that leads to wellknown aberrations in single cell or tissue imaging. Optical diffraction tomography (ODT) measures the biophysical property that causes these aberrations, the refractive index (RI). ODT is complementary to fluorescence imaging and does not require any markers. The present study introduces RI and fluorescence tomography with optofluidic rotation (RAFTOR) of suspended cells, quantifying the intracellular RI distribution and colocalizing it with fluorescence in 3D. The technique is validated with cell phantoms and used to confirm a lower nuclear RI for HL60 cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is observed in the RI distribution. The applications shown confirm predictions of previous studies and illustrate the potential of RAFTOR to improve our understanding of cells and tissues.

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Nanoscopy of bacterial cells immobilized by holographic optical tweezers

Cited by 69 publications

References 34 publications

Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V

Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V

Optical manipulation from the microscale to the nanoscale: fundamentals, advances and prospects

Three‐dimensional correlative single‐cell imaging utilizing fluorescence and refractive index tomography

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