Biology subjects

Buglakova, E.

Publications and source records attributed to Buglakova, E..

4 recordsLinked to original sources

bioRxiv2026

Automated cryo-volume EM for high-resolution 3D imaging and in situ structural analysis of cells and tissues

Cryo-volume electron microscopy (CVEM) enables three-dimensional imaging of biological ultrastructure in a near-native state but has been limited by low image contrast and charging artifacts that hinder data interpretation and complicate automation of data acquisition. Here we present an experimental and computational workflow that combines orthogonal cryo-SEM imaging, spot-geometry optimized O+ plasma-FIB milling, dedicated acquisition-control routines, and dedicated image alignment procedure. The workflow enables autonomous acquisition of volumetric datasets from vitrified cells and tissues at [~]15-20 nm isotropic resolution. In addition, sub-volume averaging of 113 nuclear pore complexes extracted from CVEM dataset of Cos-7 cell yielded its reconstruction at 9.4 nm resolution. Together, these results establish CVEM as a robust platform for autonomous high-resolution volumetric imaging and structural analysis of vitrified biological specimens.

Cell Biology↗

bioRxiv2025

A whole-body cell type atlas mapped into an electron microscopy volume of an annelid worm

Differential gene expression establishes the distinct physiology and morphology of cell types in an animal body. Single-cell sequencing and volume EM represent milestones toward the characterization of cell types, yet are difficult to combine for a comprehensive view on the cellular genotype-phenotype link. Here, we map a whole-body single-cell transcriptome into the PlatyBrowser, a multimodal cellular atlas for the marine annelid Platynereis dumerilii, and establish this combination uniquely for an entire animal. We learn that, in the 6-days-old worm, the majority of genes are tightly co-regulated to jointly implement one of eight major cellular morphotypes representing epidermis, gut, vasculature, myofibres, glia, motile cilia, glands, or neurons. Focusing on neurons, we uncover 14 families that by transcription factor identity, axonal projection, or sensory-secretory apparatus resemble conserved neuron types found in vertebrates, insects, or nematodes. We hypothesize that these existed in urbilaterian ancestors and represent the ancient core of nervous system centralization.

evolutionary biology↗

bioRxiv2023

Spatial single-cell isotope tracing reveals heterogeneity of de novo fatty acid synthesis in cancer cells

Metabolism has emerged as a key factor in homeostasis and disease including cancer. Yet, little is known about the heterogeneity of metabolic activity of cancer cells due to the lack of tools to directly probe it. Here, we present a novel method, 13C-SpaceM for spatial single-cell isotope tracing of glucose-dependent de novo lipogenesis. The method combines imaging mass spectrometry for spatially-resolved detection of 13C6-glucose-derived 13C label incorporated into esterified fatty acids with microscopy and computational methods for data integration and analysis. We validated 13C-SpaceM on a spatially-heterogeneous normoxia-hypoxia model of liver cancer cells. Investigating cultured cells, we revealed single-cell heterogeneity of lipogenic acetyl-CoA pool labelling degree upon ACLY knockdown that is hidden in the bulk analysis and its effect on synthesis of individual fatty acids. Next, we adapted 13C-SpaceM to analyze tissue sections of mice harboring isocitrate dehydrogenase (IDH)-mutant gliomas. We found a strong induction of de novo fatty acid synthesis in the tumor tissue compared to the surrounding brain. Comparison of fatty acid isotopologue patterns revealed elevated uptake of mono-unsaturated and essential fatty acids in the tumor. Furthermore, our analysis uncovered substantial spatial heterogeneity in the labelling of the lipogenic acetyl-CoA pool indicative of metabolic reprogramming during microenvironmental adaptation. Overall, 13C-SpaceM enables novel ways for spatial probing of metabolic activity at the single cell level. Additionally, this methodology provides unprecedented insight into fatty acid uptake, synthesis and modification in normal and cancerous tissues.

biochemistry↗

bioRxiv2020

Accurate 4Pi single-molecule localization using an experimental PSF model

Interferometric single-molecule localization microscopy (iPALM, 4Pi-SMS) uses multiphase interferometry to localize single fluorophores and achieves nanometer isotropic resolution in 3D. The current data analysis workflow, however, fails to reach the theoretical resolution limit due to the suboptimal localization algorithm. Here, we develop a method to fit an experimentally derived point spread function (PSF) model to the interference 4Pi-PSF. As the interference phase is not fixed with respect to the shape of the PSF, we decoupled the phase term in the model from the 3D position of the PSF. The fitter can reliably infer the interference period even without introducing astigmatism, reducing the complexity of the microscope. Using a spline-interpolated experimental PSF model and by fitting all phase images globally, we show on simulated data that we can achieve the theoretical limit of 3D resolution, the Cramer-Rao lower bound (CRLB), also for the 4Pi microscope.

biophysics↗