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Single-Molecule Fluorescence Correlation Spectroscopy (FCS)

Single-Molecule Fluorescence Correlation Spectroscopy (FCS) - BioPhysics Forum

Single-Molecule Fluorescence Correlation Spectroscopy (FCS) - Ask Questions and Discuss BioPhysics. Talk about BioPhysics Protocols and Techniques. BioPhysics Forum


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Old 04-05-2008, 05:13 AM
Pipette Filler
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Default Single-Molecule Fluorescence Correlation Spectroscopy (FCS)



Hi everyone. Here is a brief overview of Fluorescence Correlation Spectroscopy (FCS), a biophysical technique used to measure molecular interactions:

What Does FCS Measure?
Fluorescence correlation spectroscopy (FCS) is a single-molecule detection technique that detects and correlates fluctuations in fluorescence intensity within a very small detection volume (on the order of femtoliters).

What Information Do FCS Data Provide?

Concentration
Because the fluorescence intensity inside the femtoliter detection volume is directly related to the number of fluorescent particles, FCS provides a sensitive measure of concentration down to picoMolar values.

Molecular Size

By correlating the rate of fluctuations in the concentration within the detection volume, FCS also provides a measure of the rate of diffusion of particles in and out of the detection volume, which is directly related to the size of the particles.

What are the Applications of FCS?

By measuring changes in concentration and size, it's possible to monitor interactions and complexing between macromolecules such as proteins and polynucleic acids on the single-molecule scale. Basically, many of the things that you would use PCR or ELISA to detect, FCS can detect without the need for washing or signal amplification.

More Information

Here is a site that has more information on FCS theory and applications: fcsxpert.com/classroom
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Old 10-18-2010, 09:21 AM
Pipette Filler
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Default Re: Single-Molecule Fluorescence Correlation Spectroscopy (FCS)

Hello, I just started to work with FCS, I have several questions:

1. In which case it is better to use 40x water objective, and when it is better to use oil objective?
2. Triplet state and Pure diffusion fit. Which one is better to use, and why? And what is the difference?
3. Diffusion in polymers, what is the purpose of studying diffusion in polymers?

I would be very pleased if you can answer this questions in details.
Thanks in Advance.
Casper
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