Characterization of the fluorescence correlation spectroscopy (FCS) standard Rhodamine 6G and calibration of its diffusion coefficient in aqueous solutions

2014 ◽  
Vol 140 (9) ◽  
pp. 094201 ◽  
Author(s):  
G. Majer ◽  
J. P. Melchior
Pteridines ◽  
2001 ◽  
Vol 12 (4) ◽  
pp. 147-153 ◽  
Author(s):  
U. Demel ◽  
Z. Foldes-Papp ◽  
D. Fuchs ◽  
G. P. Tilz

Abstract In the present investigation, fluorescence con-elation spectroscopy (FCS) was used to measure the molecular motion of the pteridine derivative neopterin. However, technical limitations in the present optical setup precluded the identification of ,single neopterin molecules. FCS measurements with a fluorophore were also can-ied out for comparison. Exemplified by rhodamine green, we have introduced a concept that allows the detection, identification and analysis of assays in solution at the single-molecule level in tenns of bulk concentration. This concept is based on FCS and Poisson distribution analysis of assay sensitivity. The molecules had not to be quantified in a more concentrated fonn, or in flow and trapping experiments. The study demonstrated an ultrasensitive, reliable, rapid and direct tool for analytics and diagnostics in solution. We discuss a possible application of our new concept in activation control of cell-mediated immunity via neopterin determination.


ACS Nano ◽  
2015 ◽  
Vol 9 (9) ◽  
pp. 9158-9166 ◽  
Author(s):  
Lydia Kisley ◽  
Rachel Brunetti ◽  
Lawrence J. Tauzin ◽  
Bo Shuang ◽  
Xiyu Yi ◽  
...  

2004 ◽  
Vol 385 (1) ◽  
pp. 87-93 ◽  
Author(s):  
J. Toivola ◽  
P. O. Michel ◽  
L. Gilbert ◽  
T. Lahtinen ◽  
V. Marjomäki ◽  
...  

AbstractFluorescence correlation spectroscopy (FCS) was used in monitoring human parvovirus B19 virus-like particle (VLP) antibody complexes from acute phase and pastimmunity serum samples. The Oregon Green 488-labeled VLPs gave an average diffusion coefficient of 1.7x10exp-7 cm(2)s(-1) with an apparent hydrodynamic radius of 14 nm. After incubation of the fluorescent VLPs with an acute phase serum sample, the mobility information obtained from the fluorescence intensity fluctuation by autocorrelation analysis showed an average diffusion coefficient of 1.5x10exp-8 cm(2)s(-1), corresponding to an average radius of 157 nm. In contrast, incubation of the fluorescent VLPs with a pastimmunity serum sample gave an average diffusion coefficient of 3.5x10exp-8 cm(2)s(-1) and a radius of 69 nm. A control serum devoid of B19 antibodies caused a change in the diffusion coefficient from 1.7x10exp-7 to 1.6x10exp-7 cm(2)s(-1), which is much smaller than that observed with acute phase or pastimmunity sera. Thus, VLP-antibody complexes with different diffusion coefficients could be identified for the acute phase and pastimmunity sera. FCS measurement of VLPimmune complexes could be useful in distinguishing between antibodies present in acute phase or past-immunity sera as well as in titration of the VLPs.


2021 ◽  
Vol 9 ◽  
Author(s):  
Lan Yu ◽  
Yunze Lei ◽  
Ying Ma ◽  
Min Liu ◽  
Juanjuan Zheng ◽  
...  

Fluorescence correlation spectroscopy (FCS) is a powerful technique for quantification of molecular dynamics, and it has been widely applied in diverse fields, e.g., biomedicine, biophysics, and chemistry. By time-correlation of the fluorescence fluctuations induced by molecules diffusing through a focused light, FCS can quantitatively evaluate the concentration, diffusion coefficient, and interaction of the molecules in vitro or in vivo. In this review, the basic principle and implementation of FCS are introduced. Then, the advances of FCS variants are reviewed, covering dual-color FCCS, multi-focus FCS, pair correlation function (pCF), scanning FCS, focus-reduced FCS, SPIM-FCS, and inverse-FCS. Besides, the applications of FCS are demonstrated with the measurement of local concentration, hydrodynamic radius, diffusion coefficient, and the interaction of different molecules. Lastly, a discussion is given by summarizing the pros and cons of different FCS techniques, as well as the outlooks and perspectives of FCS.


2012 ◽  
Vol 117 (1) ◽  
pp. 27-33 ◽  
Author(s):  
Hyo-Sup Shin ◽  
Akimitsu Okamoto ◽  
Yasushi Sako ◽  
Sok Won Kim ◽  
Soo Yong Kim ◽  
...  

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