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Single-molecule narrowfield microscopy is a versatile tool to investigate a diverse range of protein dynamics in live cells and has been extensively used in bacteria. Here, we describe how these methods can be extended to larger eukaryotic,…

Subcellular Processes · Quantitative Biology 2016-06-09 Adam J. M. Wollman , Mark C. Leake

Super-resolution microscopy has catalyzed valuable insights into the sub-cellular, mechanistic details of many different biological processes across a wide range of cell types. Fluorescence polarization spectroscopy tools have also enabled…

Biological Physics · Physics 2023-01-25 Jack W Shepherd , Alex L Payne-Dwyer , Ji-Eun Lee , Aisha Syeda , Mark C Leake

Understanding and extracting the patterns of microscopy images has been a major challenge in the biomedical field. Although trained scientists can locate the proteins of interest within a human cell, this procedure is not efficient and…

Computer Vision and Pattern Recognition · Computer Science 2020-06-09 Hanke Chen

Human physiology and pathology arise from the coordinated interactions of diverse single cells. However, analyzing single cells has been limited by the low sensitivity and throughput of analytical methods. DNA sequencing has recently made…

Quantitative Methods · Quantitative Biology 2020-06-23 Nikolai Slavov

Super-resolution imaging techniques have largely improved our capabilities to visualize nanometric structures in biological systems. Their application further enables one to potentially quantitate relevant parameters to determine the…

Biological Physics · Physics 2019-12-18 Tina Kosǔta , Marta Cullell-Dalmau , Francesca Cella Zanacchi , Carlo Manzo

Diffusion coefficient measurements are important for many biological and material investigations, such as particle dynamics, kinetics, and size determinations. Amongst current measurement methods, single particle tracking (SPT) offers the…

Biomolecules · Quantitative Biology 2011-07-12 Shannon Kian Zareh , Michael C. DeSantis , Jonathan Kessler , Je-Luen Li , Y. M. Wang

Cellular heterogeneity is important to biological processes, including cancer and development. However, proteome heterogeneity is largely unexplored because of the limitations of existing methods for quantifying protein levels in single…

Genomics · Quantitative Biology 2018-10-29 Bogdan Budnik , Ezra Levy , Guillaume Harmange , Nikolai Slavov

Understanding sub-cellular protein localisation is an essential component to analyse context specific protein function. Recent advances in quantitative mass-spectrometry (MS) have led to high resolution mapping of thousands of proteins to…

Applications · Statistics 2019-03-12 Oliver M. Crook , Kathryn S. Lilley , Laurent Gatto , Paul D. W. Kirk

Proteins play a pivotal role in biological systems. The use of machine learning algorithms for protein classification can assist and even guide biological experiments, offering crucial insights for biotechnological applications. We…

Quantitative Methods · Quantitative Biology 2024-10-24 Yizheng Wang , Yixiao Zhai , Yijie Ding , Quan Zou

The movement of molecules inside living cells is a fundamental feature of biological processes. The ability to both observe and analyse the details of molecular diffusion in vivo at the single molecule and single cell level can add…

Quantitative Methods · Quantitative Biology 2012-11-21 Alex Robson , Kevin Burrage , Mark Leake

DNA-interacting proteins have roles multiple processes, many operating as molecular machines which undergo dynamic metastable transitions to bring about their biological function. To fully understand this molecular heterogeneity, DNA and…

Biomolecules · Quantitative Biology 2015-04-15 Adam J. M. Wollman , Helen Miller , Zhaokun Zhou , Mark C. Leake

We combine stroboscopic laser excitation with stochastic photoactivation and super-resolution fluorescence imaging. This makes it possible to record hundreds of diffusion trajectories of small protein molecules in single bacterial cells…

Quantitative Methods · Quantitative Biology 2010-03-11 Brian P. English , Arash Sanamrad , Stoyan Tankov , Vasili Hauryliuk , Johan Elf

Recent technological advances in cutting-edge ultrasensitive fluorescence microscopy have allowed single-molecule imaging experiments in living cells across all three domains of life to become commonplace. Single-molecule live-cell data is…

Biomolecules · Quantitative Biology 2015-04-15 Mark Leake

Individual cells exhibit substantial heterogeneity in protein abundance and activity, which is frequently reflected in broad distributions of fluorescently labeled reporters. Since all cellular components are intrinsically fluorescent to…

Quantitative Methods · Quantitative Biology 2023-06-13 Gabriel Torregrosa , David Oriola , Vikas Trivedi , Jordi Garcia-Ojalvo

Many DNA sequence variants influence phenotypes by altering gene expression. Our understanding of these variants is limited by sample sizes of current studies and by measurements of mRNA rather than protein abundance. We developed a…

Genomics · Quantitative Biology 2015-06-16 Frank W. Albert , Sebastian Treusch , Arthur H. Shockley , Joshua S. Bloom , Leonid Kruglyak

Here we describe the coupled standardisation of two complementary fluorescence imaging techniques and apply it to liquid-liquid phase separated condensates formed from an EGFP fluorescent reporter of Flowering Control Locus A (FCA), a…

Biomolecules · Quantitative Biology 2023-01-25 Alex L. Payne-Dwyer , Mark C. Leake

We propose a 3D convolutional neural network to simultaneously segment and detect cell nuclei in confocal microscopy images. Mirroring the co-dependency of these tasks, our proposed model consists of two serial components: the first part…

Image and Video Processing · Electrical Eng. & Systems 2018-09-07 Sundaresh Ram , Vicky T. Nguyen , Kirsten H. Limesand , Mert R. Sabuncu

Automated cell detection and localization from microscopy images are significant tasks in biomedical research and clinical practice. In this paper, we design a new cell detection and localization algorithm that combines deep convolutional…

Computer Vision and Pattern Recognition · Computer Science 2018-10-09 Yao Xue , Gilbert Bigras , Judith Hugh , Nilanjan Ray

Resolving the structural variability of proteins is often key to understanding the structure-function relationship of those macromolecular machines. Single particle analysis using Cryogenic electron microscopy (CryoEM), combined with…

Quantitative Methods · Quantitative Biology 2022-11-22 Muyuan Chen , Bogdan Toader , Roy Lederman

Multidimensional NMR spectroscopy is one of the basic tools for determining the structure of biomolecules. Unfortunately, the resolution of the spectra is often limited by inter-nuclear couplings. This limitation cannot be overcome by…

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