scholarly journals Stimulated emission depletion-based raster image correlation spectroscopy reveals biomolecular dynamics in live cells

2013 ◽  
Vol 4 (1) ◽  
Author(s):  
Per Niklas Hedde ◽  
René M. Dörlich ◽  
Rosmarie Blomley ◽  
Dietmar Gradl ◽  
Emmanuel Oppong ◽  
...  
Keyword(s):  
Correlation Spectroscopy ◽  
Stimulated Emission ◽  
Image Correlation ◽  
Live Cells ◽  
Raster Image ◽  
Biomolecular Dynamics ◽  
Image Correlation Spectroscopy ◽  
Stimulated Emission Depletion

scholarly journals Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells

2015 ◽  
Vol 2 (6) ◽  
pp. 140454 ◽  
Author(s):  
D. J. Nieves ◽  
Y. Li ◽  
D. G. Fernig ◽  
R. Lévy
Keyword(s):  
Gold Nanoparticles ◽  
Single Molecule ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Live Cells ◽  
Glycerol Water ◽  
Raster Image ◽  
Diffusion Speed ◽  
Image Correlation Spectroscopy ◽  
Representative Area

Raster image correlation spectroscopy (RICS) measures the diffusion of fluorescently labelled molecules from stacks of confocal microscopy images by analysing correlations within the image. RICS enables the observation of a greater and, thus, more representative area of a biological system as compared to other single molecule approaches. Photothermal microscopy of gold nanoparticles allows long-term imaging of the same labelled molecules without photobleaching. Here, we implement RICS analysis on a photothermal microscope. The imaging of single gold nanoparticles at pixel dwell times short enough for RICS (60 μs) with a piezo-driven photothermal heterodyne microscope is demonstrated (photothermal raster image correlation spectroscopy, PhRICS). As a proof of principle, PhRICS is used to measure the diffusion coefficient of gold nanoparticles in glycerol : water solutions. The diffusion coefficients of the nanoparticles measured by PhRICS are consistent with their size, determined by transmission electron microscopy. PhRICS was then used to probe the diffusion speed of gold nanoparticle-labelled fibroblast growth factor 2 (FGF2) bound to heparan sulfate in the pericellular matrix of live fibroblast cells. The data are consistent with previous single nanoparticle tracking studies of the diffusion of FGF2 on these cells. Importantly, the data reveal faster FGF2 movement, previously inaccessible by photothermal tracking, and suggest that inhomogeneity in the distribution of bound FGF2 is dynamic.

scholarly journals Lateral diffusion of CD14 and TLR2 in macrophage plasma membrane assessed by raster image correlation spectroscopy and single particle tracking

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sara Makaremi ◽  
Markus Rose ◽  
Suman Ranjit ◽  
Michelle A. Digman ◽  
Dawn M. E. Bowdish ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Particle Tracking ◽  
Diffusion Coefficients ◽  
Basal Membrane ◽  
Single Particle Tracking ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Live Cells ◽  
Raster Image ◽  
Image Correlation Spectroscopy

Abstract The diffusion of membrane receptors is central to many biological processes, such as signal transduction, molecule translocation, and ion transport, among others; consequently, several advanced fluorescence microscopy techniques have been developed to measure membrane receptor mobility within live cells. The membrane-anchored receptor cluster of differentiation 14 (CD14) and the transmembrane toll-like receptor 2 (TLR2) are important receptors in the plasma membrane of macrophages that activate the intracellular signaling cascade in response to pathogenic stimuli. The aim of the present work was to compare the diffusion coefficients of CD14 and TLR2 on the apical and basal membranes of macrophages using two fluorescence-based methods: raster image correlation spectroscopy (RICS) and single particle tracking (SPT). In the basal membrane, the diffusion coefficients obtained from SPT and RICS were found to be comparable and revealed significantly faster diffusion of CD14 compared with TLR2. In addition, RICS showed that the diffusion of both receptors was significantly faster in the apical membrane than in the basal membrane, suggesting diffusion hindrance by the adhesion of the cells to the substrate. This finding highlights the importance of selecting the appropriate membrane (i.e., basal or apical) and corresponding method when measuring receptor diffusion in live cells. Accurately knowing the diffusion coefficient of two macrophage receptors involved in the response to pathogen insults will facilitate the study of changes that occur in signaling in these cells as a result of aging and disease.

scholarly journals Raster image correlation spectroscopy in live cells

2010 ◽  
Vol 5 (11) ◽  
pp. 1761-1774 ◽  
Author(s):  
Molly J Rossow ◽  
Jennifer M Sasaki ◽  
Michelle A Digman ◽  
Enrico Gratton
Keyword(s):  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Live Cells ◽  
Raster Image ◽  
Image Correlation Spectroscopy

Raster image correlation spectroscopy as a novel tool to study interactions of macromolecules with nanofiber scaffolds

2011 ◽  
Vol 7 (12) ◽  
pp. 4195-4203 ◽  
Author(s):  
S.C.P. Norris ◽  
J. Humpolíčková ◽  
E. Amler ◽  
M. Huranová ◽  
M. Buzgo ◽  
...  
Keyword(s):  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Raster Image ◽  
Nanofiber Scaffolds ◽  
Image Correlation Spectroscopy

scholarly journals Measuring Lateral Diffusion of Receptors On Plasma Membrane of Macrophages Using Raster Image Correlation Spectroscopy

2018 ◽  
Vol 24 (S1) ◽  
pp. 1356-1357
Author(s):  
S. Makaremi ◽  
S. Ranjit ◽  
M.A. Digman ◽  
E. Gratton ◽  
D. M.E. Bowdish ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lateral Diffusion ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Raster Image ◽  
Image Correlation Spectroscopy

scholarly journals A nu-space for image correlation spectroscopy: characterization and application to measure protein transport in live cells

2013 ◽  
Vol 15 (8) ◽  
pp. 085006 ◽  
Author(s):  
Laurent Potvin-Trottier ◽  
Lingfeng Chen ◽  
Alan Rick Horwitz ◽  
Paul W Wiseman
Keyword(s):  
Protein Transport ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Live Cells ◽  
Image Correlation Spectroscopy

scholarly journals Measurement of Protein 53 Diffusion Coefficient in Live HeLa Cells Using Raster Image Correlation Spectroscopy (RICS)

2010 ◽  
Vol 01 (01) ◽  
pp. 31-36 ◽  
Author(s):  
Sungmin Hong ◽  
Ying-Nai Wang ◽  
Hirohito Yamaguchi ◽  
Harinibytaraya Sreenivasappa ◽  
Chao-Kai Chou ◽  
...  
Keyword(s):  
Diffusion Coefficient ◽  
Hela Cells ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Raster Image ◽  
Image Correlation Spectroscopy

scholarly journals Raster Image Correlation Spectroscopy Performance Evaluation

2019 ◽  
Vol 117 (10) ◽  
pp. 1900-1914 ◽  
Author(s):  
Marco Longfils ◽  
Nick Smisdom ◽  
Marcel Ameloot ◽  
Mats Rudemo ◽  
Veerle Lemmens ◽  
...  
Keyword(s):  
Performance Evaluation ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Raster Image ◽  
Image Correlation Spectroscopy

Dynamic range and background filtering in raster image correlation spectroscopy

2020 ◽  
Vol 279 (2) ◽  
pp. 123-138
Author(s):  
R. DE METS ◽  
A. DELON ◽  
M. BALLAND ◽  
O. DESTAING ◽  
I. WANG
Keyword(s):  
Dynamic Range ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Raster Image ◽  
Image Correlation Spectroscopy
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