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The pH of a solution containing an acid can be determined using its acid dissociation constant and its initial concentration. If a solution contains two different acids, then its pH can be determined using one of several methods depending upon the relative strength of the acids and their dissociation constants.
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Many common substances around us exist as a solution, such as ocean water, air, and gasoline. All solutions are mixtures of substances that are composed of varying amounts of two or more types of atoms or molecules. A mixture with a non-uniform composition is a heterogeneous mixture, whereas a mixture with a uniform composition is a homogeneous mixture. The components that make the homogeneous mixture are evenly spread out and thoroughly mixed. 
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Resolving Mixtures in Solution by Single-Molecule Rotational Diffusivity.

Hsiang-Yu Yang1, W E Moerner1

  • 1Department of Chemistry , Stanford University , Stanford , California 94305-4401 , United States.

Nano Letters
|July 13, 2018
PubMed
Summary
This summary is machine-generated.

This study introduces a new method to measure single-molecule rotational diffusivity, crucial for understanding biomolecular interactions. The technique accurately senses molecular size in solution without surface attachment, advancing biophysical studies.

Keywords:
ABEL trapSingle moleculefluorescence anisotropy decaymolecular sizerotational diffusion

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Area of Science:

  • Biophysics
  • Single-molecule analysis
  • Molecular dynamics

Background:

  • Sensing molecular size is vital for studying biomolecular interactions and processes like association-dissociation.
  • Rotational diffusivity is more sensitive to molecular size and conformation than translational diffusivity.
  • Existing single-molecule rotational diffusivity measurements face limitations like low photon counts and perturbative surface attachment.

Purpose of the Study:

  • To develop and demonstrate a novel method for measuring single-molecule rotational diffusivity in solution.
  • To overcome the limitations of existing techniques, enabling accurate size sensing without surface attachment.
  • To simultaneously determine multiple properties of single molecules, including rotational diffusivity and fluorescence characteristics.

Main Methods:

  • Combining the solution-phase Anti-Brownian ELectrokinetic (ABEL) trap with maximum likelihood analysis of time-resolved fluorescence anisotropy.
  • Utilizing the information inherent in each detected photon for precise measurements.
  • Analyzing time resolution and precision through statistical signal analysis and simulations.

Main Results:

  • Successfully resolved a mixture of single- and double-stranded DNA molecules based on their rotational diffusivity.
  • Simultaneously determined rotational diffusivity, fluorescence brightness, lifetime, and anisotropy for individual DNA molecules.
  • Identified key parameters for selecting fluorescent labels to optimize molecular size information extraction.

Conclusions:

  • The developed method enables accurate measurement of single-molecule rotational diffusivity in native solution environments.
  • This technique provides a powerful, non-perturbative tool for investigating biomolecular interactions and size.
  • The findings offer guidance for selecting appropriate fluorescent labels for advanced single-molecule biophysical studies.