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Open AccessJournal ArticleDOI

A practical guide to single-molecule FRET

TLDR
A practical guide to using smFRET, focusing on the study of immobilized molecules that allow measurements of single-molecule reaction trajectories from 1 ms to many minutes, is provided.
Abstract
Single-molecule fluorescence resonance energy transfer (smFRET) is one of the most general and adaptable single-molecule techniques. Despite the explosive growth in the application of smFRET to answer biological questions in the last decade, the technique has been practiced mostly by biophysicists. We provide a practical guide to using smFRET, focusing on the study of immobilized molecules that allow measurements of single-molecule reaction trajectories from 1 ms to many minutes. We discuss issues a biologist must consider to conduct successful smFRET experiments, including experimental design, sample preparation, single-molecule detection and data analysis. We also describe how a smFRET-capable instrument can be built at a reasonable cost with off-the-shelf components and operated reliably using well-established protocols and freely available software.

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Journal ArticleDOI

Fluorescence Lifetime Measurements and Biological Imaging

TL;DR: The lifetime of a photophysical process is the time required by a population of N electronically excited molecules to be reduced by a factor of e via the loss of energy through fluorescence and other non-radiative processes and the average length of time τ is called the mean lifetime, or simply lifetime.
Journal ArticleDOI

Properties and Applications of Colloidal Nonspherical Noble Metal Nanoparticles

TL;DR: This Review highlights morphology-dependent properties of nonspherical noble metal nanoparticles with a focus on localized surface plasmon resonance and local field enhancement, as well as their applications in various fields including Raman spectroscopy, fluorescence enhancement, analytics and sensing, photothermal therapy, (bio-)diagnostics, and imaging.
Journal ArticleDOI

Nanoantenna-enhanced gas sensing in a single tailored nanofocus

TL;DR: This work places a single palladium nanoparticle near the tip region of a gold nanoantenna and detects the changing optical properties of the system upon hydrogen exposure, demonstrating antenna-enhanced hydrogen sensing at the single-particle level.

Methods and Applications

TL;DR: The aim of the research presented in this thesis is to create new methods for design for manufacturing, by using several approaches of KE, and find the beneficial and less beneficial aspects of these methods in comparison to each other and earlier research.
Journal ArticleDOI

SLC6 Neurotransmitter Transporters: Structure, Function, and Regulation

TL;DR: The understanding of the molecular structure, function, and pharmacology of these proteins has advanced rapidly, and intensive efforts have been directed toward understanding the molecular and cellular mechanisms involved in regulation of the activity of this important class of transporters, leading to new methodological developments and important insights.
References
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Book

A Laboratory manual

M. Ashburner
TL;DR: To develop a program to print the barcodes using two commonly uses command sets and hence evaluates their ease of use for such applications, students should be able to program dot matrix printers, by manipulating bit level information and ink jet printers using page description language, such as PCL.
Journal ArticleDOI

Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).

TL;DR: A high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores that can, in principle, reach molecular-scale resolution is developed.
BookDOI

Single-channel recording

Bert Sakmann, +1 more
TL;DR: A Practical Guide to Patch Clamping R.H. Penner, C.W. Heinemann, and P. Jonas.
Journal ArticleDOI

A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions

TL;DR: These procedures, which can circumvent the need for large-scale phage or plasmid growths, preparative gel-electrophoresis and the screening of molecular clones, can facilitate the rapid study of sequence-specific interactions of proteins and DNA.
Journal ArticleDOI

Fluorescence spectroscopy of single biomolecules.

TL;DR: The progress in applying single-molecule detection and single-Molecule spectroscopy at room temperature by laser-induced fluorescence with the use of fluorophores that are site-specifically attached to macromolecules is reviewed.
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