A method for single molecule tracking using a conventional single-focus confocal setup

Sina Jazani, Ioannis Sgouralis, Steve Presse

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

One way to achieve spatial resolution using fluorescence imaging - and track single molecules - is to use wide-field illumination and collect measurements over multiple sensors (camera pixels). Here we propose another way that uses confocal measurements and a single sensor. Traditionally, confocal microscopy has been used to achieve high temporal resolution at the expense of spatial resolution. This is because it utilizes very few, and commonly just one, sensors to collect data. Yet confocal data encode spatial information. Here we show that non-uniformities in the shape of the confocal excitation volume can be exploited to achieve spatial resolution. To achieve this, we formulate a specialized hidden Markov model and adapt a forward filtering-backward sampling Markov chain Monte Carlo scheme to efficiently handle molecular motion within a symmetric confocal volume characteristically used in fluorescence correlation spectroscopy. Our method can be used for single confocal volume applications or incorporated into larger computational schemes for specialized, multi-confocal volume, optical setups.

Original languageEnglish (US)
Article number114108
JournalJournal of Chemical Physics
Volume150
Issue number11
DOIs
StatePublished - Mar 21 2019

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Molecules
spatial resolution
Sensors
Fluorescence
molecules
sensors
Confocal microscopy
Optical resolving power
Hidden Markov models
fluorescence
Markov processes
Markov chains
Lighting
Pixels
Cameras
temporal resolution
Spectroscopy
nonuniformity
Sampling
Imaging techniques

ASJC Scopus subject areas

  • Physics and Astronomy(all)
  • Physical and Theoretical Chemistry

Cite this

A method for single molecule tracking using a conventional single-focus confocal setup. / Jazani, Sina; Sgouralis, Ioannis; Presse, Steve.

In: Journal of Chemical Physics, Vol. 150, No. 11, 114108, 21.03.2019.

Research output: Contribution to journalArticle

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