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DNA Nanotechnology pp 273–289Cite as

Optimal Practices for Surface-Tethered Single Molecule Total Internal Reflection Fluorescence Resonance Energy Transfer Analysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 749))

Abstract

Single molecule fluorescence microscopy can be used to follow the mechanics of molecular biology processes in real time. However, many factors, from flow cell preparation to improper data analysis can negatively impact single molecule fluorescence resonance energy transfer (smFRET) experiments. Here, we describe some best practices for ensuring that smFRET data are of the highest quality. In addition to instrumentation, we describe sample preparation and data analysis.

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Acknowledgments

This work was supported by RO1 GM077872 (S.H.L.) from the National Institutes of Health.

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Authors and Affiliations

  1. Departments of Cell Biology and Physiology and Bioengineering, University of Pittsburgh School of Medicine and Swanson School of Engineering, Petersen Institute of NanoScience and Engineering, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    Sanford H. Leuba

Authors
  1. Matt V. Fagerburg
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  2. Sanford H. Leuba
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Corresponding author

Correspondence to Sanford H. Leuba .

Editor information

Editors and Affiliations

  1. Dipto. Biochimica, Università Bologna, Via Irnerio 48, Bologna, 40126, Italy

    Giampaolo Zuccheri

  2. Dipto. Biochimica, Università Bologna, Via Irnerio 48, Bologna, 40126, Italy

    Bruno Samorì

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Fagerburg, M.V., Leuba, S.H. (2011). Optimal Practices for Surface-Tethered Single Molecule Total Internal Reflection Fluorescence Resonance Energy Transfer Analysis. In: Zuccheri, G., Samorì, B. (eds) DNA Nanotechnology. Methods in Molecular Biology, vol 749. Humana Press. https://doi.org/10.1007/978-1-61779-142-0_19

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  • DOI: https://doi.org/10.1007/978-1-61779-142-0_19

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-141-3

  • Online ISBN: 978-1-61779-142-0

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