Mastering
single-molecule FRET:
A practical handbook for researchers

Understand the fundamentals of smFRET and explore its potential in your own research​

Discover everything you need to understand single-molecule FRET and use it to advance your research

This practical smFRET handbook brings together the core principles, experimental workflows, and analytical strategies that underpin modern single-molecule Förster Resonance Energy Transfer (smFRET). Designed for researchers at every stage – from newcomers to seasoned practitioners – it distils the essential knowledge required to generate high-quality smFRET data and interpret it with confidence.

Inside, you’ll find:

  • Clear, accessible explanations of key concepts such as FRET efficiency, stoichiometry, ALEX, burst analysis, and accurate FRET correction

  • Step-by-step descriptions of how a confocal smFRET instrument works, how single molecules are detected, and how to ensure true single-molecule sensitivity in practice

  • Illustrative examples and case studies showing how smFRET resolves structural heterogeneity, conformational dynamics, and nanoscale distance changes in biomolecules

  • Comparisons to complementary single-molecule and structural biology techniques, helping you understand when – and why – smFRET is the method of choice

  • Insights into powerful analytical approaches, including E–S plotting, burst search strategies, and correction-factor estimation for high-precision distance measurements

Download this guide if you want to:

introduce a simple, solution-based method into your existing workflows that rapidly resolves conformational changes at the single molecule level

complement existing static workflows with dynamic data

 unlock deeper functional insights and novel biological questions

smFRET explained in 4 minutes

Watch the essentials before downloading the handbook 👇

What questions does the handbook answer?

  • What is FRET?
  • What are smFRET measurements?
  • What is FRET efficiency and stoichiometry?
  • What is Alternating Laser EXcitation (ALEX)?
  • How does a confocal smFRET instrument work?
  • What does smFRET data look like?
  • What are the key considerations for researchers?
  • What are the applications of smFRET?
  • What is accurate FRET correction and why does it matter?

Want to know more about bringing dynamic single-molecule techniques to your benchtop?

We believe in making innovative techniques, such as freely diffusing smFRET, accessible to all researchers, regardless of their experience or infrastructure. 

Our benchtop tools:

  • Are incredibly easy to use – no intensive training or years of expertise required, no complicated sample prep or instrument alignment needed, and results produced within minutes
  • Are compact enough to use on your bench top, in ambient light – no darkroom, optical table or laser safe room required, fitting easily into your current infrastructure
  • Offer simple workflows that can be implemented with minimal changes to existing workflows
  • Come with highly easy-to-use acquisition and analysis software that combines over 50 years of experience, bringing a ‘professor in a box’ into your lab
  • Publication-ready data in minutes
EI-FLEX pro instrument for single-molecule FRET (smFRET)