scholarly journals A simulation-guided fluorescence correlation spectroscopy tool to investigate the protonation dynamics of cytochrome c oxidase

2016 ◽  
Vol 18 (18) ◽  
pp. 12877-12885 ◽  
Author(s):  
A. Wolf ◽  
C. Schneider ◽  
T.-Y. Kim ◽  
K. Kirchberg ◽  
P. Volz ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation ◽  
Rate Dependent ◽  
Confocal Volume ◽  
Dependent Processes

Fluorescence correlation spectroscopy (FCS) is a single molecule based technique to temporally resolve rate-dependent processes by correlating the fluorescence fluctuations of individual molecules traversing through a confocal volume.

scholarly journals A fluorescence correlation spectrometer for measurements in cuvettes

2018 ◽  
Author(s):  
B Sahoo ◽  
TB Sil ◽  
B Karmakar ◽  
K Garai
Keyword(s):  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Diffusion Time ◽  
Correlation Spectroscopy ◽  
Ease Of Use ◽  
Fluorescence Correlation ◽  
Wide Range ◽  
Correlation Spectrometer ◽  
Confocal Volume ◽  
Working Distance

ABSTRACTWe have developed a fluorescence correlation spectroscopy (FCS) setup for performing single molecule measurements on samples inside regular cuvettes. We built this by using an Extra Long Working Distance (ELWD), 0.7 NA, air objective with working distance > 1.8 mm. We have achieved counts per molecule > 44 kHz, diffusion time < 64 μs for rhodamine B in aqueous buffer and a confocal volume < 2 fl. The cuvette-FCS can be used for measurements over a wide range of temperature that is beyond the range permitted in the microscope-based FCS. Finally, we demonstrate that cuvette-FCS can be coupled to automatic titrators to study urea dependent unfolding of proteins with unprecedented accuracy. The ease of use and compatibility with various accessories will enable applications of cuvette-FCS in the experiments that are regularly performed in fluorimeters but are generally avoided in microscope-based FCS.

Fluorescence correlation spectroscopy: The technique and its applications in soft matter

2018 ◽  
Vol 4 (4) ◽  
Author(s):  
Anjali Gupta ◽  
Jagadish Sankaran ◽  
Thorsten Wohland
Keyword(s):  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Soft Matter ◽  
Photophysical Properties ◽  
Spatiotemporal Analysis ◽  
Correlation Spectroscopy ◽  
Basic Principles ◽  
Chemical Reaction Kinetics ◽  
Fluorescence Correlation ◽  
Wide Range

Abstract Fluorescence correlation spectroscopy (FCS) is a well-established single-molecule method used for the quantitative spatiotemporal analysis of dynamic processes in a wide range of samples. It possesses single-molecule sensitivity but provides ensemble averaged molecular parameters such as mobility, concentration, chemical reaction kinetics, photophysical properties and interaction properties. These parameters have been utilized to characterize a variety of soft matter systems. This review provides an overview of the basic principles of various FCS modalities, their instrumentation, data analysis, and the applications of FCS to soft matter systems.

Characterization of Pteridines: a New Approach by Fluorescence Correlation Spectroscopy and Analysis of Assay Sensitivity

Pteridines ◽  
2001 ◽  
Vol 12 (4) ◽  
pp. 147-153 ◽  
Author(s):  
U. Demel ◽  
Z. Foldes-Papp ◽  
D. Fuchs ◽  
G. P. Tilz
Keyword(s):  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Correlation Spectroscopy ◽  
Cell Mediated Immunity ◽  
Distribution Analysis ◽  
Assay Sensitivity ◽  
New Approach ◽  
Single Molecule Level ◽  
Fluorescence Correlation

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.

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