Three-dimensional biplane spectroscopic single-molecule localization microscopy

Song, Ki Hee; Zhang, Yang; Wang, Gaoxiang; Sun, Cheng

doi:10.1364/optica.6.000709

Cited by 34 publications

(42 citation statements)

References 32 publications

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“…We characterized the spectroscopic signatures of nanospheres using the spectral centroid method 5,13 . Figure 2g shows the scatter plot of the photon count versus spectral centroid for five nanospheres.…”

Section: Single and Multicolour Sdssmlm Imaging Of Nanospheresmentioning

confidence: 99%

“…The ability of spectroscopic single-molecule localization microscopy (sSMLM) to capture the spectroscopic signatures of individual molecules along with their spatial distribution allows the observation of subcellular structures and dynamics at the nanoscale. As a result, sSMLM has shown great potential in understanding fundamental biomolecular processes in cell biology and material science [1][2][3][4][5][6][7] . It also enables the characterization of nanoparticle properties based on the emission spectrum at the single-particle level [8][9][10] .…”

Section: Introductionmentioning

confidence: 99%

“…Similar to other localizationbased super-resolution techniques, such as stochastic optical reconstruction microscopy (STORM) and point accumulation for imaging in nanoscale topography (PAINT), the localization precision of sSMLM is fundamentally limited by the number of collected photons per emitter 11 . However, sSMLM suffers from further photon budget constraints since the collected photons of each molecule need to be divided into two separate channels to simultaneously capture the spatial and spectral information [5][6][7][8][9][10] . Thus, the spatial localization precision of sSMLM also depends on the splitting ratio between the spatial and spectral channels and is typically limited to 15-30 nm in cell imaging 2,3,5,6 .…”

Section: Introductionmentioning

confidence: 99%

“…However, sSMLM suffers from further photon budget constraints since the collected photons of each molecule need to be divided into two separate channels to simultaneously capture the spatial and spectral information [5][6][7][8][9][10] . Thus, the spatial localization precision of sSMLM also depends on the splitting ratio between the spatial and spectral channels and is typically limited to 15-30 nm in cell imaging 2,3,5,6 . Although a dual-objective sSMLM design was previously demonstrated with improved spatial localization precision, it imposes a constraint on live-cell imaging and adds complexity to system alignment 1 .…”

Section: Introductionmentioning

confidence: 99%

“…Although a dual-objective sSMLM design was previously demonstrated with improved spatial localization precision, it imposes a constraint on live-cell imaging and adds complexity to system alignment 1 . The splitting of photons into two channels in sSMLM forces an inherent trade-off between the spatial and spectral localization precisions 5 . Currently, a method to fully utilize the full photon budget to maximize both the spatial and spectral localization precisions in sSMLM is lacking.…”

Section: Introductionmentioning

confidence: 99%

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Symmetrically dispersed spectroscopic single-molecule localization microscopy

et al. 2020

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Spectroscopic single-molecule localization microscopy (sSMLM) was used to achieve simultaneous imaging and spectral analysis of single molecules for the first time. Current sSMLM fundamentally suffers from a reduced photon budget because the photons from individual stochastic emissions are divided into spatial and spectral channels. Therefore, both spatial localization and spectral analysis only use a portion of the total photons, leading to reduced precisions in both channels. To improve the spatial and spectral precisions, we present symmetrically dispersed sSMLM, or SDsSMLM, to fully utilize all photons from individual stochastic emissions in both spatial and spectral channels. SDsSMLM achieved 10-nm spatial and 0.8-nm spectral precisions at a total photon budget of 1000. Compared with the existing sSMLM using a 1:3 splitting ratio between spatial and spectral channels, SDsSMLM improved the spatial and spectral precisions by 42% and 10%, respectively, under the same photon budget. We also demonstrated multicolour imaging of fixed cells and three-dimensional single-particle tracking using SDsSMLM. SDsSMLM enables more precise spectroscopic single-molecule analysis in broader cell biology and material science applications.

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Section: Single and Multicolour Sdssmlm Imaging Of Nanospheresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Symmetrically dispersed spectroscopic single-molecule localization microscopy

et al. 2020

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Lokaler Wassergehalt in Polymergelen bestimmt über superauflösende Fluoreszenzlebensdauer‐Bildgebung

Jana,

Nevskyi,

Höche

et al. 2024

Angewandte Chemie

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Water molecules play an important role in the structure, function, and dynamics of (bio‐)materials. A direct access to the number of water molecules in nanoscopic volumes can thus give new molecular insights into materials and allow for fine‐tuning their properties in sophisticated applications. The determination of the local water content has become possible by the finding that H2O quenches the fluorescence of red‐emitting dyes. Since deuterated water, D2O, does not induce significant fluorescence quenching, fluorescence lifetime measurements performed in different H2O/D2O‐ratios yield the local water concentration. We combined this effect with the recently developed fluorescence lifetime single molecule localization microscopy imaging (FL‐SMLM) in order to nanoscopically determine the local water concentration in microgels, i.e. soft hydrogel particles consisting of a cross‐linked polymer swollen in water. The change in water content of thermo‐responsive microgels when changing from their swollen state at room temperature to a collapsed state at elevated temperature could be analyzed. A clear decrease in water content was found that was, to our surprise, rather uniform throughout the entire microgel volume. Only a slightly higher water concentration around the dye was found in the periphery with respect to the center of the swollen microgels.

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