Rapid ensemble measurement of protein diffusion and probe blinking dynamics in cells

Sehayek, Simon; Yi, Xiyu; Weiss, Shimon; Wiseman, Paul W.

doi:10.1016/j.bpr.2021.100015

Cited by 2 publications

(5 citation statements)

References 57 publications

(78 reference statements)

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“…The kICS has been previously developed by Kolin et al In this work, an extension kICS analysis was performed as has previously been described in Sehayek et al In brief, in this extension, the autocorrelation considers intensity fluctuations over time, δ t , as shown in eq . normalΦ̃ ( k , τ ) = ⟨ δ t ι̃ false( bold-italick , t false) · δ t ι * ∼ false( bold-italick , t + τ false) ⟩ t …”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…Data Analysis. The determination of diffusion coefficients by extended kICS and iMSD was performed in MATLAB using an algorithm developed by Sehayek et al; 18 the expanded kICS was analyzed in ROIs of 40 × 40 pixels toward the edge of the cell. QD trajectories were analyzed using ImageJ particle tracker plug-in to get the particle positions.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…Later, they developed an extension of kICS that, unlike the original method, included parameters dealing with fluorophore photophysics such as fluorescence bleaching and blinking. 18 This kICS extension has been successfully used for measuring lateral dynamics in real heterogeneities in space and time in living cells. 7,17,19,20 An alternative strategy based on ICS offers a comprehensive analysis of the full space−time correlation function (CF) by introducing the spatiotemporal image correlation spectroscopy (STICS) method.…”

mentioning

confidence: 99%

“…Wiseman et al developed a novel approach, known as k -space image correlation spectroscopy (kICS), for measuring the dynamics and number densities of fluorescently tagged membrane proteins. Later, they developed an extension of kICS that, unlike the original method, included parameters dealing with fluorophore photophysics such as fluorescence bleaching and blinking . This kICS extension has been successfully used for measuring lateral dynamics in real heterogeneities in space and time in living cells. ,,, …”

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confidence: 99%

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Angiotensin II Treatment Induces Reorganization and Changes in the Lateral Dynamics of Angiotensin II Type 1 Receptor in the Plasma Membrane Elucidated by Photoactivated Localization Microscopy Combined with Image Spatial Correlation Analysis

et al. 2022

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The mechanisms by which angiotensin II type 1 receptor is distributed and the diffusional pattern in the plasma membrane (PM) remain unclear, despite their crucial role in cardiovascular homeostasis. In this work, we obtained quantitative information of angiotensin II type 1 receptor (AT1R) lateral dynamics as well as changes in the diffusion properties after stimulation with ligands in living cells using photoactivated localization microscopy (PALM) combined with image spatial–temporal correlation analysis. To study the organization of the receptor at the nanoscale, expansion microscopy (ExM) combined with PALM was performed. This study revealed that AT1R lateral diffusion increased after binding to angiotensin II (Ang II) and the receptor diffusion was transiently confined in the PM. In addition, ExM revealed that AT1R formed nanoclusters at the PM and the cluster size significantly decreased after Ang II treatment. Taking these results together suggest that Ang II binding and activation cause reorganization and changes in the dynamics of AT1R at the PM.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: ■ Experimental Sectionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Angiotensin II Treatment Induces Reorganization and Changes in the Lateral Dynamics of Angiotensin II Type 1 Receptor in the Plasma Membrane Elucidated by Photoactivated Localization Microscopy Combined with Image Spatial Correlation Analysis

et al. 2022

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Recent Advances in Fluorescence Recovery after Photobleaching for Decoupling Transport and Kinetics of Biomacromolecules in Cellular Physiology

et al. 2022

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Among the new molecular tools available to scientists and engineers, some of the most useful include fluorescently tagged biomolecules. Tools, such as green fluorescence protein (GFP), have been applied to perform semi-quantitative studies on biological signal transduction and cellular structural dynamics involved in the physiology of healthy and disease states. Such studies focus on drug pharmacokinetics, receptor-mediated endocytosis, nuclear mechanobiology, viral infections, and cancer metastasis. In 1976, fluorescence recovery after photobleaching (FRAP), which involves the monitoring of fluorescence emission recovery within a photobleached spot, was developed. FRAP allowed investigators to probe two-dimensional (2D) diffusion of fluorescently-labelled biomolecules. Since then, FRAP has been refined through the advancements of optics, charged-coupled-device (CCD) cameras, confocal microscopes, and molecular probes. FRAP is now a highly quantitative tool used for transport and kinetic studies in the cytosol, organelles, and membrane of a cell. In this work, the authors intend to provide a review of recent advances in FRAP. The authors include epifluorescence spot FRAP, total internal reflection (TIR)/FRAP, and confocal microscope-based FRAP. The underlying mathematical models are also described. Finally, our understanding of coupled transport and kinetics as determined by FRAP will be discussed and the potential for future advances suggested.

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Rapid ensemble measurement of protein diffusion and probe blinking dynamics in cells

Cited by 2 publications

References 57 publications

Angiotensin II Treatment Induces Reorganization and Changes in the Lateral Dynamics of Angiotensin II Type 1 Receptor in the Plasma Membrane Elucidated by Photoactivated Localization Microscopy Combined with Image Spatial Correlation Analysis

Angiotensin II Treatment Induces Reorganization and Changes in the Lateral Dynamics of Angiotensin II Type 1 Receptor in the Plasma Membrane Elucidated by Photoactivated Localization Microscopy Combined with Image Spatial Correlation Analysis

Recent Advances in Fluorescence Recovery after Photobleaching for Decoupling Transport and Kinetics of Biomacromolecules in Cellular Physiology

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