Photophysical processes in single molecule organic fluorescent probes

Elana M S Stennett, Monika A. Ciuba, Marcia Levitus

Research output: Contribution to journalReview articlepeer-review

226 Scopus citations


The use of organic fluorescent probes in biochemical and biophysical applications of single molecule spectroscopy and fluorescence microscopy techniques continues to increase. As single molecule measurements become more quantitative and new developments in super-resolution imaging allow researchers to image biological materials with unprecedented resolution, it is becoming increasingly important to understand how the properties of the probes influence the signals measured in these experiments. In this review, we focus on the photochemical and photophysical processes of organic fluorophores that affect the properties of fluorescence emission. This includes photobleaching, quenching, and the formation of non-emissive (dark) states that result in fluorescence blinking in a variety of timescales. These processes, if overlooked, can result in an erroneous interpretation of the data. Understanding their physical origins, on the other hand, allows researchers to design experiments and interpret results so that the maximum amount of information about the system of interest can be extracted from fluorescence signals.

Original languageEnglish (US)
Pages (from-to)1057-1075
Number of pages19
JournalChemical Society Reviews
Issue number4
StatePublished - Feb 21 2014

ASJC Scopus subject areas

  • Chemistry(all)


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