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research article

Resolution of fluorescence correlation measurements

Meseth, Ulrich
•
Wohland, Thorsten
•
Rigler, Rudolf
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1999
Biophysical Journal

The resoln. limit of fluorescence correlation spectroscopy for two-component solns. is investigated theor. and exptl. The autocorrelation function for two different particles in soln. were computed, statistical noise was added, and the resulting curve was fitted with a least squares fit. These simulations show that the ability to distinguish between two different mol. species in soln. depends strongly on the no. of photons detected from each particle, their difference in size, and the concn. of each component in soln. To distinguish two components, their diffusion times must differ by at least a factor of 1.6 for comparable quantum yields and a high fluorescence signal. Expts. were conducted with Rhodamine 6G and Rhodamine-labeled bovine serum albumin. The exptl. results support the simulations. In addn., they show that even with a high fluorescence signal but significantly different quantum yields, the diffusion times must differ by a factor much bigger than 1.6 to distinguish the two components. Depending on the quantum yields and the difference in size, there exists a concn. threshold for the less abundant component below which it is not possible to det. with statistical means alone that two particles are in soln. [on SciFinder (R)]

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Type
research article
DOI
10.1016/S0006-3495(99)77321-2
Web of Science ID

WOS:000078972300042

Author(s)
Meseth, Ulrich
•
Wohland, Thorsten
•
Rigler, Rudolf
•
Vogel, Horst  
Date Issued

1999

Published in
Biophysical Journal
Volume

76

Issue

3

Start page

1619

End page

1631

Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LCPPM  
Available on Infoscience
February 27, 2006
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/226330
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