Abstract
One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.
| Original language | English |
|---|---|
| Title of host publication | Single Molecule Analysis |
| Subtitle of host publication | Methods and Protocols |
| Publisher | Humana Press Inc |
| Pages | 93-113 |
| Number of pages | 21 |
| ISBN (Electronic) | 9781493972715 |
| DOIs | |
| Publication status | Published - 2018 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1665 |
| ISSN (Print) | 1064-3745 |
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Keywords
- Confocal fluorescence
- Fluorophore
- Microscopy
- TIRF
- Wide-field epi-fluorescence
Cite this
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A brief introduction to single-molecule fluorescence methods. / van den Wildenberg, Siet M.J.L.; Prevo, Bram; Peterman, Erwin J.G.
Single Molecule Analysis: Methods and Protocols. Humana Press Inc, 2018. p. 93-113 (Methods in Molecular Biology; Vol. 1665).Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic
TY - CHAP
T1 - A brief introduction to single-molecule fluorescence methods
AU - van den Wildenberg, Siet M.J.L.
AU - Prevo, Bram
AU - Peterman, Erwin J.G.
PY - 2018
Y1 - 2018
N2 - One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.
AB - One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.
KW - Confocal fluorescence
KW - Fluorophore
KW - Microscopy
KW - TIRF
KW - Wide-field epi-fluorescence
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U2 - 10.1007/978-1-4939-7271-5_5
DO - 10.1007/978-1-4939-7271-5_5
M3 - Chapter
T3 - Methods in Molecular Biology
SP - 93
EP - 113
BT - Single Molecule Analysis
PB - Humana Press Inc
ER -