scholarly journals Characterizing diffusion dynamics of a membrane protein associated with nanolipoproteins using fluorescence correlation spectroscopy

2011 ◽  
Vol 20 (2) ◽  
pp. 437-447 ◽  
Author(s):  
Tingjuan Gao ◽  
Craig D. Blanchette ◽  
Wei He ◽  
Feliza Bourguet ◽  
Sonny Ly ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Fluorescence Correlation Spectroscopy ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation ◽  
Diffusion Dynamics

scholarly journals Advanced processing and analysis of conventional confocal microscopy generated scanning FCS data

2017 ◽  
Author(s):  
Dominic Waithe ◽  
Falk Schneider ◽  
Jakub Chojnacki ◽  
Mathias Clausen ◽  
Dilip Shrestha ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Spatial Heterogeneity ◽  
Fluorescence Correlation Spectroscopy ◽  
Large Scale ◽  
Molecular Diffusion ◽  
Correlation Spectroscopy ◽  
The Other ◽  
Statistical Accuracy ◽  
Fluorescence Correlation ◽  
Diffusion Dynamics

AbstractScanning Fluorescence Correlation Spectroscopy (scanning FCS) is a variant of conventional point FCS that allows molecular diffusion at multiple locations to be measured simultaneously. It enables disclosure of potential spatial heterogeneity in molecular diffusion dynamics and also the acquisition of a large amount of FCS data at the same time, providing large statistical accuracy. Here, we optimize the processing and analysis of these large-scale acquired sets of FCS data. On one hand we present FoCuS-scan, scanning FCS software that provides an end-to-end solution for processing and analysing scanning data acquired on commercial turnkey confocal systems. On the other hand, we provide a thorough characterisation of large-scale scanning FCS data over its intended time-scales and applications and propose a unique solution for the bias and variance observed when studying slowly diffusing species. Our manuscript enables researchers to straightforwardly utilise scanning FCS as a powerful technique for measuring diffusion across a broad range of physiologically relevant length scales without specialised hardware or expensive software.

scholarly journals Quantifying Interactions of a Membrane Protein Embedded in a Lipid Nanodisc using Fluorescence Correlation Spectroscopy

2014 ◽  
Vol 106 (2) ◽  
pp. L05-L08 ◽  
Author(s):  
Sonny Ly ◽  
Feliza Bourguet ◽  
Nicholas O. Fischer ◽  
Edmond Y. Lau ◽  
Matthew A. Coleman ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Fluorescence Correlation Spectroscopy ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation

CHARACTERIZATION OF POLY(ACRYLIC ACID) DIFFUSION DYNAMICS ON THE GRAFTED SURFACE OF POLY(ETHYLENE TEREPHTHALATE) FILMS BY FLUORESCENCE CORRELATION SPECTROSCOPY

2006 ◽  
Vol 01 (04) ◽  
pp. 433-441 ◽  
Author(s):  
XIAOTAO PAN ◽  
CAILI AW ◽  
YANAN DU ◽  
HANRY YU ◽  
THORSTEN WOHLAND
Keyword(s):  
Ionic Strength ◽  
Acrylic Acid ◽  
Chain Length ◽  
Fluorescence Correlation Spectroscopy ◽  
Ethylene Terephthalate ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation ◽  
Poly Ethylene Terephthalate ◽  
Diffusion Dynamics ◽  
Poly Ethylene

Poly(acrylic acid) (PAAc) is a commonly used polymer grafted on poly(ethylene terephthalate) films for the immobilization of bioactive molecules that directly interact with living cells or tissues for the maintenance of their viability and functionality. The diffusion property of the grafted PAAc on the surface is a critical parameter related to the density, length of polymer chains, and ionic strength of the solution. Fluorescence correlation spectroscopy (FCS) is able to measure the diffusion coefficient of fluorescent particles in solution with single molecule sensitivity and specificity. It was used as an effective tool to detect diffusion dynamics of Atto 565 molecules, a good indicator for viscosity of PAAc, in both aqueous polymer solutions and polymer grafted film surfaces immersed in solution. In this work we determine the polymer chain length under different polymerization conditions in solution and deduce the solution viscosity by FCS measurements of Atto 565 as tracer molecule. By using the same tracer on the grafted polymer films we can infer the viscosity of these grafted layers under a variety of conditions, including the PAAc chain length, the UV exposure time during polymerization, the ionic strength, and the pH value of the immersed solution.

scholarly journals Fluorescence Correlation Spectroscopy Measurement Based on Fiber Optics for Biological Materials

2021 ◽  
Vol 11 (15) ◽  
pp. 6744
Author(s):  
Johtaro Yamamoto ◽  
Akira Sasaki
Keyword(s):  
Fiber Optics ◽  
Fluorescence Correlation Spectroscopy ◽  
Fiber Optic ◽  
Fluorescent Dyes ◽  
Water Immersion ◽  
Correlation Spectroscopy ◽  
Objective Lens ◽  
Fluorescence Correlation ◽  
Diffusion Dynamics ◽  
Fluorescent Molecules

A robust fluorescence correlation spectroscopy system called fiber-optic based fluorescence correlation spectroscopy (FB-FCS) was developed; this system enables the measurement of diffusion dynamics and concentration of fluorescent molecules based on the principle of fluorescence correlation spectroscopy without any mechanical adjustment of the experimental setup. The system consisted of fiber optics and a water-immersion objective lens. The hydrodynamic diameters and concentrations of organic fluorescent dyes and fluorescently labeled proteins were successfully measured. Because of the fiber-optic-based setup, the FB-FCS system is compact and inexpensive. We expect FB-FCS to be suitable for use in laboratories, medical diagnosis, and environmental measurements.

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