A5013316072- Advanced Fluorescence Microscopy Techniques
- Genomics and Chromatin Dynamics
- Advanced Electron Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Near-Field Optical Microscopy
- Epigenetics and DNA Methylation
- Single-cell and spatial transcriptomics
- Gene expression and cancer classification
- RNA and protein synthesis mechanisms
- Photoacoustic and Ultrasonic Imaging
- DNA and Nucleic Acid Chemistry
- Immune cells in cancer
- RNA Interference and Gene Delivery
- RNA Research and Splicing
- Spectroscopy Techniques in Biomedical and Chemical Research
- Optical Coherence Tomography Applications
- Protein Degradation and Inhibitors
- Advanced X-ray Imaging Techniques
- Neuroinflammation and Neurodegeneration Mechanisms
- Identification and Quantification in Food
- Face and Expression Recognition
- Brain Tumor Detection and Classification
- DNA Repair Mechanisms
- Digestive system and related health
- Integrated Circuits and Semiconductor Failure Analysis
Saveetha University
2025
Institute of Cancer Research
2022-2024
Edinburgh Cancer Research
2024
Delft University of Technology
2024
Royal Marsden NHS Foundation Trust
2023
National Physical Laboratory
2017-2022
University of Cambridge
2017-2022
Wellcome/MRC Cambridge Stem Cell Institute
2018-2021
Carnegie Institution for Science
2017-2021
Department of Embryology
2017-2021
Super-resolution microscopy (SRM) is a fast-developing field that encompasses fluorescence imaging techniques with the capability to resolve objects below classical diffraction limit of optical resolution. Acknowledged Nobel prize in 2014, numerous SRM methods have meanwhile evolved and are being widely applied biomedical research, all specific strengths shortcomings. While some capable nanometre-scale molecular resolution, others geared towards volumetric three-dimensional multi-colour or...
Light microscopy enables researchers to observe cellular mechanisms with high spatial and temporal resolution. However, the increasing complexity of current imaging technologies, coupled financial constraints potential users, hampers general accessibility reach cutting-edge microscopy. Open can address this issue by making well-designed well-documented hardware software solutions openly available a broad audience. In Comment, we provide definition open present recent projects in field. We...
Several approaches have been described to fluorescently label and image DNA chromatin in situ on the single-molecule level. These superresolution microscopy techniques are based detecting optically isolated, tagged anti-histone antibodies, labeled precursor analogs, or fluorescent dyes bound DNA. Presently they suffer from various drawbacks such as low labeling efficiency interference with structure. In this report, we demonstrate that minor groove binding dyes, Hoechst 33258, 33342, DAPI,...
Significance The structure of the pachytene chromosome is essential to understand how genetic recombination can happen during meiosis. Using single molecule localization microscopy with DNA staining, we show that chromatin heavily constrained by defined periodic clusters along synaptonemal complex (SC). Staining various posttranslational histone modifications further reveals associated three distinct nanoscale compartments. Whereas active transcription emanates both axially and radially in...
Cells detect and adapt to hypoxic nutritional stress through immediate transcriptional, translational metabolic responses. The environmental effects of ischemia on chromatin nanostructure were investigated using single molecule localization microscopy DNA binding dyes acetylated histones, by the sensitivity digestion with DNAseI, fluorescence recovery after photobleaching (FRAP) core linker histones. Short-term oxygen nutrient deprivation cardiomyocyte cell line HL-1 induces a previously...
Structured illumination microscopy (SIM) has emerged as an essential technique for three-dimensional (3D) and live-cell super-resolution imaging. However, to date, there not been a dedicated workshop or journal issue covering the various aspects of SIM, from bespoke hardware software development use commercial instruments biological applications. This special aims recap recent developments well outline future trends. In addition we cover related topics such complementary techniques,...
MINFLUX is purported as the next revolutionary fluorescence microscopy technique claiming a spatial resolution in range of 1-3 nm fixed and living cells. Though claim molecular attractive, I am concerned whether true 1 has been attained. Here, compare performance with other super-resolution methods focusing particularly on claims, subjective filtering localizations, detection versus labelling efficiency possible limitations when imaging biological samples containing densely labelled...
Histone modifications alone or in combination are thought to modulate chromatin structure and function; a concept termed histone code. By combining evidence from several studies, we investigated if the code can play role higher-order folding of chromatin. Firstly using genomic data, analyzed associations between at nucleosome level. We could dissect composition individual nucleosomes into five predicted clusters modifications. Secondly, by assembling raw reads various length scales, noticed...
Single Molecule Localization Microscopy (SMLM) is a recently emerged optical imaging method that was shown to achieve resolution in the order of tens nanometers intact cells. Novel high methods might be crucial for understanding how chromatin, complex DNA and proteins, arranged eukaryotic cell nucleus. Such an approach utilizing switching fluorescent, DNA-binding dye Vybrant® DyeCycle™ Violet has been previously demonstrated by us (Żurek-Biesiada et al., 2015) [1]. Here we provide...
We report that high-density single-molecule super-resolution microscopy can be achieved with a conventional epifluorescence microscope set-up and mercury arc lamp. The configuration termed as laser-free (LFSM) is an extension of localization (SMLM) techniques allows single molecules to switched on off (a phenomenon ‘blinking’), detected localized. use short burst deep blue excitation (350–380 nm) further used reactivate the blinking, once blinking process has slowed or stopped. A resolution...
Abstract During organogenesis, patterns and gradients of gene expression underlie organization diversified cell specification to generate complex tissue architecture. While the cerebral cortex is organized into six excitatory neuronal layers, it unclear whether glial cells are mimic laminae or show distinct layering. To determine molecular architecture mammalian cortex, we developed a high content pipeline that can quantify single-cell in situ . The Large-area Spatial Transcriptomic (LaST)...
In single-molecule microscopy, a big question is how precisely we can estimate the location of single molecule. Our research shows that by using iterative localisation microscopy and factoring in prior information, boost precision reduce number photons needed. Leveraging Van Trees inequality aids determining optimal achievable. approach holds promise for wider application discerning across diverse imaging scenarios, encompassing various illumination strategies, point spread functions...
MINFLUX is a promising new development in single-molecule localization microscopy, claiming resolution of 1-3 nm living and fixed biological specimens. While can achieve very high localisation precision, quantitative analysis reported results leads us to dispute the claim question reliability for imaging sub-100-nm structural features, its current state.
During meiosis, homologous chromosomes associate to form a unique structure called synaptonemal complex (SC) whose disruption leads infertility. Information about the epigenetic features of chromatin within this at level super-resolution microscopy is largely lacking. We combined single molecule localization with quantitative analytical methods describe landscape meiotic pachytene stage in mouse oocytes. DNA found be non-randomly distributed along length SC condensed clusters. Periodic...
Histones are punctuated with small chemical modifications that alter their interaction DNA. One attractive hypothesis stipulates certain combinations of these histone may function, alone or together, as a part predictive code to provide ground rules for chromatin folding. We consider four features relate folding: charge neutralisation, molecular specificity, robustness and evolvability. Next, we present evidence the association among different at various levels organisation show how...
Gwosch et al. (2020) and Balzarotti (2017) purport MINFLUX as the next revolutionary fluorescence microscopy technique claiming a spatial resolution in range of 1-3 nm fixed living cells. Though claim molecular is attractive, I am concerned whether true 1 has been attained. Here, compare performance with other super-resolution methods focussing particularly on claims, atypical image rendering, visualisation enhancement, subjective filtering localizations, detection vs labelling efficiency...