bioRxiv Science⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.1101/752287

Sub-Nanometer Precision using Bayesian Grouping of Localizations

Fazel, M.·Wester, M. J.·Rieger, B.·Jungmann, R.·Lidke, K. A.
View original source ↗Export metadata ↓

Abstract

Single-molecule localization microscopy super-resolution methods such as DNA-PAINT and (d)STORM can generate multiple observed localizations over the time course of data acquisition from each dye or binding site that are not a priori assigned to those specific dyes or binding sites. We describe a Bayesian method of grouping and combining localizations from multiple blinking/binding events that can improve localization precision to better than one nanometer. The known statistical distribution of the number of binding/blinking events per dye/docking strand along with the precision of each localization event are used to estimate the true number and location of emitters in closely-spaced clusters.

Explore related subjects

biophysicsPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Fazel, M., Wester, M. J., Rieger, B., Jungmann, R., Lidke, K. A.. 2019-08-30. Sub-Nanometer Precision using Bayesian Grouping of Localizations. https://doi.org/10.1101/752287

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Kinetic gating by an iron sulfur chain modulates flavin radical-pair spin chemistry in mitochondrial Complex I

Mitochondrial Complex I generates reactive oxygen species at its flavin cofactor, where spin dependent reaction pathways have been proposed. Models of this chemistry treat the flavin oxygen radical pair in isolation, although the flavin is wired to a chain of eight iron sulfur clusters. Here we model the pair with this chain using the human Complex I structure and open quantum system models of spin coupling and electron transfer. In the simulated network, spin coherence decays rapidly along the chain, whereas proximal clusters alter the calculated radical-pair singlet yield. Their influence is kinetically gated: a downstream cluster affects the modelled yield when its electron-transfer rate competes with radical pair recombination, a threshold robust to driving force and to the transfer model. Modelling does set the sign: one directional spin hops predict suppression and inflate coherence transport, whereas charge resolved transfer drives the singlet yield towards its spin-randomized limit unless triplet recombination dominates. Detectable spin dependent effects in this framework require superoxide spin relaxation to be substantially slower than in free solution, a condition not yet established for Complex I.

biophysics↗
bioRxiv2026

Semidilute Polymer Scaling Controls PEG-induced Protein Phase Separation

When does polyethylene glycol (PEG) mass concentration provide a common measure of crowding strength across polymer chain lengths? Here we use bovine serum albumin (BSA) and PEG as a controlled depletion system and measure both coexisting phases to determine binodals, tie lines, and critical points across PEG molecular weights of 4--100 kDa. Phase diagrams for PEG 20, 35, and 100 kDa approximately collapse in mass-concentration coordinates, consistent with semidilute polymer scaling. Generalized free volume theory (GFVT) describes the BSA binodals and tie lines. It also approximately describes binodals for three additional globular proteins in PEG 35K, extending the comparison across proteins spanning 7.2--66.4 kDa. For the binary PEG mixtures tested, GFVT approximates the binodals using an effective polymer size corresponding to the weight-average molecular weight. These results identify a regime in which PEG mass concentration (g/L) provides a transferable measure of depletion-induced crowding.

biophysics↗
bioRxiv2026

Correlation-resolved sum-frequency generation microscopy of biological tissue with bright twin beams

Sum-frequency generation (SFG) driven by bright twin beams generated through high-gain parametric down-conversion comprises a coherent component arising from correlated mode pairs and an incoherent component arising from interactions between uncorrelated modes. Modal correlations can enhance coherent SFG in anisotropic nonlinear media, for example by extending the effective interaction length. Whether such correlations can likewise benefit image formation in heterogeneous biological tissue remains unknown. Here we address this question by performing SFG imaging of biological tissue using bright twin beams as the excitation source. Spectral decomposition of the SFG signal separates its coherent and incoherent components, allowing us to reconstruct co-registered images and examine how twin-beam correlations influence image formation. In mouse skeletal muscle, the coherent-component image revealed sarcomere periodicity with a Bragg-spot-to-background ratio of approximately 310, compared with 7.5 in the incoherent-component image. In fresh human dermis, the coherent-component image showed greater Fourier power than the incoherent-component image in the 0.30-0.50 \mu m^{-1} band and a greater fractional autocovariance decay at short spatial lags. These results demonstrate that twin-beam correlations can enhance contrast and detectability at selected spatial frequencies in heterogeneous tissue. Correlation-resolved SFG microscopy thus offers a route for exploiting bright quantum-light sources in nonlinear optical bioimaging.

biophysics↗