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Related Concept Videos

Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

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Atomic fluorescence spectroscopy (AFS) is an analytical technique that involves the electronic transitions of atoms in a flame, furnace, or plasma being excited by electromagnetic (EM) radiation. When these atoms absorb energy, they become excited and subsequently release energy as they return to their original state. This emitted light, or "fluorescence," is observed at a right angle to the incident beam. Both absorption and emission processes transpire at distinct wavelengths, which...
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Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...
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Circles01:18

Circles

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A circle in the coordinate plane is defined as the set of all points that lie at a constant distance, known as the radius, from a fixed point called the center. This relationship is captured using the distance formula. For a point (x, y) on the circle and a center (h, k), the distance between them equals the radius r. By squaring both sides of the distance formula, the equation of the circle is written in standard form:Constructing the Equation from Geometric InformationIf the center and the...
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Correlations02:20

Correlations

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Correlation means that there is a relationship between two or more variables (such as ice cream consumption and crime), but this relationship does not necessarily imply cause and effect. When two variables are correlated, it simply means that as one variable changes, so does the other. We can measure correlation by calculating a statistic known as a correlation coefficient. A correlation coefficient is a number from -1 to +1 that indicates the strength and direction of the relationship between...
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Statistical tests can calculate whether there is a relationship, or correlation, between independent and dependent variables. An indirect relationship of the variables signifies a correlation, while a direct relationship shows causation. If it is determined that no connection exists between the variables, then the correlation is a coincidence.
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2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

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Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
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Scanning fluorescence correlation spectroscopy comes full circle.

German Gunther1, David M Jameson1, Joao Aguilar1

  • 1Laboratorio de Cinética y Fotoquímica, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile; Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI, USA; Universidad de Concepción, Facultad de Ciencias Químicas, Departamento de Polímeros, Concepción, Chile.

Methods (San Diego, Calif.)
|February 7, 2018
PubMed
Summary
This summary is machine-generated.

This review covers fluorescence correlation spectroscopy (FCS) for live cell studies. Circular scanning FCS, with photon counting histogram analysis, reveals details about yeast ribosomal structures and live cell lipid domains.

Keywords:
Circular scanning FCSFCS

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Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy FCS
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Area of Science:

  • Biophysics
  • Cell Biology
  • Spectroscopy

Background:

  • Fluorescence Correlation Spectroscopy (FCS) is a powerful technique for analyzing molecular dynamics in solutions and complex biological systems.
  • Live cell imaging presents unique challenges due to cellular complexity and dynamic environments.

Purpose of the Study:

  • To provide a comprehensive review of Fluorescence Correlation Spectroscopy (FCS) applications in live cell studies.
  • To detail the principles, instrumentation, and data analysis of circular scanning FCS.
  • To illustrate the utility of FCS in investigating cellular structures and dynamics.

Main Methods:

  • Overview of the theoretical underpinnings of FCS.
  • Detailed discussion of circular scanning FCS instrumentation and data processing.
  • Application of photon counting histogram analysis with scanning FCS.
  • Utilizing dual-channel detection in conjunction with scanning FCS.

Main Results:

  • Demonstration of FCS's capability to probe yeast ribosomal structures in vivo.
  • Illustration of scanning FCS with dual-channel detection for studying live cell lipid domains.
  • Highlighting the quantitative information obtainable from FCS regarding molecular interactions and dynamics.

Conclusions:

  • Fluorescence Correlation Spectroscopy (FCS) is a versatile tool for quantitative analysis of molecular behavior in live cells.
  • Circular scanning FCS offers enhanced capabilities for studying complex cellular systems.
  • FCS methods provide valuable insights into cellular structures and dynamics, aiding in understanding cellular function.