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Multiphoton Microscopy of Entire Intact Mouse Organs

Sonia Parra, Josepfh Zinter, Michael Levene
2010 Biophysical Journal  
Using multi-color single particle tracking fluorescence microscopy, we have probed the intracellular degradation of low-density lipoprotein in living cells.  ...  Transient colocalization of LDL with specific vesicles and the intensity of the LDL particle are measured simultaneously.  ...  Using multi-color single particle tracking fluorescence microscopy, we have probed the intracellular degradation of low-density lipoprotein in living cells.  ... 
doi:10.1016/j.bpj.2009.12.3138 fatcat:aaq6xqfg7vbuxnnvyfgmj563om

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Timo Appelhans, Felix R.M. Beinlich, Christian P. Richter, Rainer Kurre, Karin B. Busch
2018 Journal of Visualized Experiments  
Moreover, by multi-color localization microscopy, the localization and tracking profiles of proteins in different subcompartments are obtained simultaneously.  ...  The technique allows for multi-color imaging of single molecules, whereby the number of colors is limited by the available membrane-permeable dyes and the repertoire of selflabeling enzymes.  ...  material, and the CellNanOs board for providing microscopes for use.  ... 
doi:10.3791/57690 pmid:30010642 pmcid:PMC6102026 fatcat:qz5u3lim2rgwbnb5qg4e767iwe

Imaging and Quantitation Techniques for Tracking Cargo along Endosome-to-Golgi Transport Pathways

Pei Chia, Paul Gleeson
2013 Cells  
Here, we review these advances, as well as highlight developments in quantitative image analysis and novel unbiased analytical approaches to quantitate protein localization.  ...  Recent improvements in the resolution of light microscopy, coupled with the development of a range of fluorescent-based probes, have provided new approaches to dissecting membrane domains and the regulation  ...  However, colocalization analysis in optical microscopy is a process that can be ambiguous and inconsistent.  ... 
doi:10.3390/cells2010105 pmid:24709647 pmcid:PMC3972656 fatcat:fmskw75l2fbd7edr3gwtbijwuq

Imaging Dopamine and Serotonin in Live Neurons with Multi-Photon Excited Ultraviolet Auto-Fluorescence

Arkarup Bandyopadhyay, Suman Nag, Manoj Kumar, Shyamala Mani, Sudipta Maiti
2010 Biophysical Journal  
Both methods allow for quantitative measurement of fluorescent particle densities and oligomerization states within individual images acquired with laser-scanning microscopy.  ...  We used fluorescent moment image analysis and spatial intensity distribution analysis (SpIDA) to study the oligomeric state of NBCe1-A in cultured cells expressing the cotransporter and in rat kidney tissue  ...  Using multi-color single particle tracking fluorescence microscopy, we have probed the intracellular degradation of low-density lipoprotein in living cells.  ... 
doi:10.1016/j.bpj.2009.12.3133 fatcat:2adxshw4n5faxjk2mi6rlinaza

Application of Super-Resolution and Advanced Quantitative Microscopy to the Spatio-Temporal Analysis of Influenza Virus Replication

Emma Touizer, Christian Sieben, Ricardo Henriques, Mark Marsh, Romain F. Laine
2021 Viruses  
In recent years, advances in light microscopy, including super-resolution microscopy and single-molecule imaging, have enabled many viral replication steps to be visualised dynamically in living cells.  ...  In this review, we discuss how state-of-the-art imaging technologies, notably quantitative live-cell and super-resolution microscopy, are providing new nanoscale and molecular insights into influenza virus  ...  In Figure 1d , fluorescence light-sheet microscopy in living cells combined with quantitative particle tracking in 3D showed the co-transport of Rab11A-containing recycling endosomes with influenza viral  ... 
doi:10.3390/v13020233 pmid:33540739 fatcat:uixg6gbyvbfingfilkodbligwa

High-Throughput Single Extracellular Vesicle Profiling [chapter]

Yanling Cai, Di Wu
2021 Secretomics [Working Title]  
In this chapter, we will discuss advanced strategies for studying single EVs, including single EV imaging systems, flow cytometers, nano-sensing technologies and single EV barcoding assay.  ...  Extracellular vesicles (EVs) are heterogeneous due to their cell of origins, biogenesis, stimuli in the microenvironment and so on. Single EV analysis is required for the study of EV heterogeneity.  ...  The colocalization of multiple biomarkers could be simultaneously recorded via multi-color TIR imaging.  ... 
doi:10.5772/intechopen.97544 fatcat:7ioe3zroqfgjrffgpiw5vdyeqy

Analytical tools for single-molecule fluorescence imaging in cellulo [article]

Mark Leake
2015 arXiv   pre-print
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  ...  robustly localize and track single fluorescently-labelled molecules, algorithms to objectively interpret molecular mobility, analysis protocols to reliably estimate molecular stoichiometry and turnover  ...  The author would particularly like to thank his students, postdocs and collaborators co-authored on many of the studies cited in this article, and to the growing collegiate fellowship of the BPSI at the  ... 
arXiv:1504.03486v1 fatcat:tqnnkzp6r5e6ler77mi5wkrw4y

Single Particle Tracking Reveals that EGFR Signaling Activity Is Amplified in Clathrin-Coated Pits

Jenny Ibach, Yvonne Radon, Márton Gelléri, Michael H. Sonntag, Luc Brunsveld, Philippe I. H. Bastiaens, Peter J. Verveer, James Keen
2015 PLoS ONE  
We employed single particle tracking to relate the mobility and aggregation of EGFR to its signaling activity. EGFR mobility alternates between short-lived free, confined and immobile states.  ...  In the immobile state, EGFR tends to aggregate in clathrin-coated pits, which is further enhanced in a phosphorylation-dependent manner and does not require ligand binding.  ...  Liguzinski and M. Dittmann for benzylguanine labeling, P. Bosch and V. Subramaniam for discussions on single particle tracking, and F. Persson and M.  ... 
doi:10.1371/journal.pone.0143162 pmid:26575183 pmcid:PMC4648588 fatcat:czrvb7cnx5ewxob6hyqn35q3my

Analytical tools for single-molecule fluorescence imaging in cellulo

M. C. Leake
2014 Physical Chemistry, Chemical Physics - PCCP  
The author would particularly like to thank his students, postdocs and collaborators co-authored on many of the studies cited in this article, and to the growing collegiate fellowship of the BPSI at the  ...  Raw data were kindly donated by Quan Xue, Oliver Harriman and Erik Hedlund. Many thanks to Nick Jones for critical discussion.  ...  Considering the particle intensity models from fluorescence microscopy, first a set of filters are applied to the image data to suppress noise and enhance the contrast of moving particles in each image  ... 
doi:10.1039/c4cp00219a pmid:24626744 fatcat:32mg4tmiire23mrh5226zqr5su

PySTACHIO: Python Single-molecule TrAcking stoiCHiometry Intensity and simulatiOn, a flexible, extensible, beginner-friendly and optimized program for analysis of single-molecule microscopy data [article]

Jack W Shepherd, Ed Higgins, Adam JM Wollman, Mark C Leake
2021 bioRxiv   pre-print
It can also evaluate diffusion coefficients for the relatively short single-particle tracking data that is characteristic of time-resolved image stacks.  ...  As highly sensitive camera pixel sensor arrays have grown both larger and faster and optical microscopy techniques become ever more refined, there has been an explosion in the quantity of data acquired  ...  Acknowledgements This work was supported by funding from the Leverhulme Trust (RPG-2019-156) and the Biotechnology and Biological Sciences Research Council BBSRC (BB/R001235/1).  ... 
doi:10.1101/2021.03.18.435952 fatcat:jvjijl4zajd67arm7x4jo6g2ba

Novel Tools to Measure Single Molecules Colocalization in Fluorescence Nanoscopy by Image Cross Correlation Spectroscopy

Simone Pelicci, Laura Furia, Mirco Scanarini, Pier Giuseppe Pelicci, Luca Lanzanò, Mario Faretta
2022 Nanomaterials  
images in correlative multiresolution, multimodal imaging.  ...  We propose new tools based on Image Cross Correlation Spectroscopy (ICCS) to quantify the colocalization of fluorescent signals at single molecule level.  ...  S.P. and M.F. want to thank Francesco Nuti for technical assistance in microscope set-up. Conflicts of Interest: The authors declare no conflict of interest.  ... 
doi:10.3390/nano12040686 pmid:35215014 pmcid:PMC8875509 fatcat:yl3vpcsehvg57bb6h34lf27e2e

Imaging Flow Cytometry and Confocal Immunofluorescence Microscopy of Virus-Host Cell Interactions

Ryley D. McClelland, Tyce N. Culp, David J. Marchant
2021 Frontiers in Cellular and Infection Microbiology  
This review will discuss how confocal microscopy and imaging flow cytometry are used to investigate virus entry and replication mechanisms in fixed and live cells.  ...  Quantification of viral images and the use of cryo-electron microscopy to gather structural information of viruses is also explored.  ...  When combined with fluorescent microscopy and supravital dyes, live-cell imaging greatly enhances our understanding of the dynamic and multi-step nature of viral infections.  ... 
doi:10.3389/fcimb.2021.749039 pmid:34712624 pmcid:PMC8546218 fatcat:5rkdv53szrfnbcj6bvoytzdxje

Applying Single Molecule Fluorescence Microscopy to Precisely Count Absolute Numbers of Proteins in Living Cells

Matthieu G.S. Palayret, Steven F. Lee, Ricardo A. Fernandes, Helena Browne, Simon J. Davis, David Klenerman
2013 Biophysical Journal  
Our findings have important implications for super-resolution microscopy, particularly in the context of quantitative imaging and co-localization in multi-color super-resolution experiments. 2696-Pos Board  ...  Then the channel was used as a template to precisely characterize photoactivation and photoconversion efficiency at the single molecule level of most commonly used fluorescent proteins in PALM microscopy  ...  -Pos Board B716 Three-Dimensional Tracking of Fluorescent Particles in Confocal Microscopy Trevor Ashley, Catherine Chan-Tse, Jeffrey Worthey, Sean Andersson.  ... 
doi:10.1016/j.bpj.2012.11.2908 fatcat:wju3ipmetfgjjbjir5nijhv3ie

Highly fluorescent biodegradable PLGA nano-carriers allows real-time tracking of individual particles in vivo [article]

Igor Khalin, Caterina Severi, Doriane Heimburger, Antonia Wehn, Farida Hellal, Andreas Reisch, Andrey Klymchenko, Nikolaus Plesnila
2020 bioRxiv   pre-print
Using a novel ex vivo imaging protocol we unambiguously distinguished nanoparticles' fluorescence from tissue auto-fluorescence and demonstrate they were taken up into late endothelial lysosomes.  ...  In summary, by increasing the brightness of clinically approved PLGA nanocarriers to a level which allowed in vivo tracking, we were able to demonstrate that PF-68 shifts the uptake of individual particles  ...  in vivo and ex vivo at single particle resolution by single-and multi-photon microscopy.  ... 
doi:10.1101/2020.11.19.385062 fatcat:l7zupph4jje33nmk4l7zzs7goi

Single Lipid Molecule Dynamics on Supported Lipid Bilayers with Membrane Curvature

Philip Cheney, Alan Weisgerber, Alec Feuerbach, Michelle Knowles
2017 Membranes  
Both lipids studied contain 16 carbon, saturated tails and a head group tag for fluorescence microscopy measurements.  ...  We then tested whether single biotinylated DHPE molecules sense curvature using single particle tracking methods.  ...  Location-Guided Colocalization Image Analysis Location-guided colocalization methods were used and these methods are visually summarized in Figure 1 .  ... 
doi:10.3390/membranes7010015 pmid:28294967 pmcid:PMC5371976 fatcat:urhc5hj7xnd3xhlfwomxvsd5aa
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