A5052240435- Advanced Fluorescence Microscopy Techniques
- Developmental Biology and Gene Regulation
- Cell Image Analysis Techniques
- Genomics and Chromatin Dynamics
- Cellular Mechanics and Interactions
- RNA Research and Splicing
- Gene expression and cancer classification
- Advanced Electron Microscopy Techniques and Applications
- Fuzzy Logic and Control Systems
- Zebrafish Biomedical Research Applications
- Advanced Biosensing Techniques and Applications
- Neural Networks and Applications
- Molecular Biology Techniques and Applications
- Near-Field Optical Microscopy
- Epigenetics and DNA Methylation
- Retinal Development and Disorders
- Congenital heart defects research
- Advanced biosensing and bioanalysis techniques
- Force Microscopy Techniques and Applications
- Photoreceptor and optogenetics research
- Chromosomal and Genetic Variations
- Optical Coherence Tomography Applications
- Photoacoustic and Ultrasonic Imaging
- Neuroscience and Neuropharmacology Research
- Vibrio bacteria research studies
University of Cambridge
2015-2024
Anglia Ruskin University
2024
Centre de Génétique Moléculaire
2014-2016
Institut de Biologie de l'École Normale Supérieure
2013-2016
École Normale Supérieure - PSL
2013-2016
Université Paris-Sud
2014
Centre National de la Recherche Scientifique
2013-2014
Johannes Kepler University of Linz
2005-2013
Laboratoire de Géologie de l’École Normale Supérieure
2012
MIT Lincoln Laboratory
1985-2005
Transcription is reported to be spatially compartmentalized in nuclear transcription factories with clusters of RNA polymerase II (Pol II). However, little known about when these foci assemble or their relative stability. We developed a quantitative single-cell approach characterize protein spatiotemporal organization, single-molecule sensitivity live eukaryotic cells. observed that Pol form transiently, an average lifetime 5.1 (± 0.4) seconds, which refutes the notion they are statically...
Abstract The molecular features of synapses in the hippocampus underpin current models learning and cognition. Although synapse ultra-structural diversity has been described canonical hippocampal circuitry, our knowledge sub-synaptic organisation synaptic molecules remains largely unknown. To address this, mice were engineered to express Post Synaptic Density 95 protein (PSD95) fused either eGFP or mEos2 imaged with two orthogonal super-resolution methods: gated stimulated emission depletion...
How genetic programs generate cell-intrinsic forces to shape embryos is actively studied, but less so how tissue-scale physical impact morphogenesis. Here we address the role of latter during axis extension, using Drosophila germband extension (GBE) as a model. We found previously that cells elongate in anteroposterior (AP) extending germband, suggesting an extrinsic tensile force contributed body extension. further characterized AP cell elongation patterns GBE, by tracking and quantifying...
Abstract The mammary gland undergoes cycles of growth and regeneration throughout reproductive life, a process that requires stem cells (MaSCs). Whilst recent genetic fate-mapping studies using lineage-specific promoters have provided valuable insights into the epithelial hierarchy, true differentiation potential adult MaSCs remains unclear. To address this, herein we utilize stochastic genetic-labelling strategy to indelibly mark single cell its progeny in situ , combined with tissue...
A key feature of Notch signaling is that it directs immediate changes in transcription via the DNA-binding factor CSL, switching from repression to activation. How generates both a sensitive and accurate response-in absence any amplification step-remains be elucidated. To address this question, we developed real-time analysis CSL dynamics including single-molecule tracking vivo. In Notch-OFF nuclei, small proportion molecules transiently binds DNA, while Notch-ON conditions recruitment...
Clonal analysis is helping us understand the dynamics of cell replacement in homeostatic adult tissues (Simons and Clevers, 2011). Such an analysis, however, has not yet been achieved for continuously growing tissues, but essential if we wish to architecture organs. The retinas lower vertebrates grow throughout life, from retinal stem cells (RSCs) progenitor (RPCs) at rim retina, called ciliary marginal zone (CMZ). Here, show that RSCs reside a niche extreme periphery CMZ divide...
During embryonic nervous system assembly, mRNA localization is precisely regulated in growing axons, affording subcellular autonomy by allowing controlled protein expression space and time. Different sets of mRNAs exhibit different patterns across the axon. However, little known about how move axons or these are generated. Here, we couple molecular beacon technology with highly inclined laminated optical sheet microscopy to image single molecules identified endogenous axons. By combining...
During gastrulation, embryonic cells become specified into distinct germ layers. In mouse, this continues throughout somitogenesis from a population of bipotent stem called neuromesodermal progenitors (NMps). However, the degree self-renewal associated with NMps in fast-developing zebrafish embryo is unclear. Using genetic clone-tracing method, we labelled early and found strong clonal similarity between spinal cord mesoderm tissues. We followed individual cell lineages using light-sheet...
The segregation of bacterial chromosomes follows a precise choreography spatial organisation. It is initiated by the bipolar migration sister copies replication origin (ori). Most contain partition system (Par) with parS sites in close proximity to ori that contribute active mobilisation region towards old pole. This thought result longitudinal chromosomal arrangement within cell. In this study, we followed duplication frequency and cellular position 19 Vibrio cholerae genome loci as...
We introduce single molecule light field microscopy (SMLFM), a new class of three-dimensional (3D) localization microscopy. By segmenting the back focal plane microscope objective with an array microlenses to generate multiple 2D perspective views, same fluorophore can be imaged from different angles. These in combination bespoke fitting algorithm, enable 3D positions fluorophores determined parallax. SMLFM achieves up 20 nm precision throughout extended <mml:math...
Abstract Some compulsive disorders have been considered to stem from the loss of control over coping strategies, such as displacement. However, cellular mechanisms involved in acquisition behaviours and their subsequent manifestation vulnerable individuals not elucidated. Considering role locus coeruleus (LC) noradrenaline-dependent system stress related excessive behaviours, we hypothesised that neuroplastic changes LC may be associated with an adjunctive polydipsic water drinking, a...
We developed a microarray platform for PCR amplification-independent expression profiling of minute samples. A novel scanning system combined with specialized biochips enables detection down to individual fluorescent oligonucleotide molecules specifically hybridized their complementary sequence over the entire biochip surface cm 2 size. limit 1.3 fM target concentration—corresponding only 39,000 in sample solution—and dynamic range 4.7 orders magnitude have been achieved. The applicability...
The replication terminus region (Ter) of the unique chromosome most bacteria locates at mid-cell time cell division. In several species, this localization participates in necessary coordination between segregation and division, notably for selection division site, licensing machinery assembly correct alignment dimer resolution sites. genome Vibrio cholerae, agent deadly human disease cholera, is divided into two chromosomes, chrI chrII. Previous fluorescent microscopy observations suggested...
The planar orientation of cell division (OCD) is important for epithelial morphogenesis and homeostasis. Here, we ask how mechanics antero-posterior (AP) patterning combine to influence the first divisions after gastrulation in Drosophila embryonic epithelium. We analyse hundreds show that stress anisotropy, notably from compressive forces, can reorient directly metaphase. Stress anisotropy influences OCD by imposing metaphase elongation, despite mitotic rounding, overrides interphase...
Epithelial folding shapes embryos and tissues during development. Here, we investigate the coupling between epithelial actomyosin-enriched compartmental boundaries. The mechanistic relationship two is unclear, because boundaries are not necessarily associated with folds. Also, some cases of occur independently actomyosin contractility. We investigated shallow folds called parasegment grooves that form at anterior posterior compartments in early
A major issue in the field of fuzzy applications is complexity algorithms used. In order to obtain efficient methods, it necessary reduce without losing easy interpretability components. One possibilities achieve reduction combine rule interpolation with use hierarchical structured bases, as proposed by Sugeno et al. (1991). For interpolation, method Koczy and Hirota (1993) used, but other techniques are also suggested. The difficulty applying this that often impossible determine a partition...
Abstract Light-sheet microscopy is an increasingly popular technique in the life sciences due to its fast 3D imaging capability of fluorescent samples with low photo toxicity compared confocal methods. In this work we present a new, fast, flexible and simple implement method optimize illumination light-sheet requirement at hand. A telescope composed two electrically tuneable lenses enables us define thickness position independently but accurately within milliseconds therefore image quality...
Abstract We study the problem of deconvolution for light-sheet microscopy, where data is corrupted by spatially varying blur and a combination Poisson Gaussian noise. The spatial variation point spread function microscope determined interaction between excitation sheet detection objective PSF. introduce model image formation process that incorporates this we formulate variational accounts noise through fidelity term consisting infimal convolution single fidelities, first introduced in L....
While the biochemistry of gene transcription has been well studied, our understanding how this process is organised in 3D within intact nucleus less understood. Here we investigate structure actively transcribed chromatin and architecture its interaction with active RNA polymerase. For analysis, have used super-resolution microscopy to image Drosophila melanogaster Y loops which represent huge, several megabases long, single units. The provide a particularly amenable model system for...
Advanced paternal age increases the risk of transmitting de novo germline mutations, particularly missense mutations activating receptor tyrosine kinase (RTK) signaling pathway, exemplified by FGFR3 mutation linked to achondroplasia (ACH). This is attributed expansion spermatogonial stem cells carrying mutation, forming sub-clonal clusters in ageing testis, thereby increasing frequency mutant sperm and number affected offspring from older fathers. While prior studies proposed a correlation...
Advanced paternal age increases the risk of transmitting de novo germline mutations, particularly missense mutations activating receptor tyrosine kinase (RTK) signalling pathway, as exemplified by
Abstract Chromatin state is thought to impart regulatory function the underlying DNA sequence. This can be established through histone modifications and chromatin organisation, but exactly how these factors relate one another regulate gene expression unclear. In this study, we have used super-resolution microscopy image Y loops of Drosophila melanogaster primary spermatocytes, which are enormous transcriptionally active fibres, each representing single transcription units that individually...