Saturation-compensated measurements for fluorescence lifetime imaging microscopy

Article

Abstract

Fluorophore saturation is the key factor limiting the speed and excitation range of fluorescence lifetime imaging microscopy (FLIM). For example, fluorophore saturation causes incorrect lifetime measurements when using conventional frequency-domain FLIM at high excitation powers. In this Letter, we present an analytical theoretical description of this error and present a method for compensating for this error in order to extract correct lifetime measurements in the limit of fluorophore saturation. We perform a series of simulations and experiments to validate our methods. The simulations and experiments show a 13.2× and a 2.6× increase in excitation range, respectively. The presented method is based on algorithms that can be easily applied to existing FLIM setups.

Attributes

Attribute NameValues
Creator
  • Genevieve D. Vigil

  • Lina Cao

  • Aamir A. Khan

  • David Benirschke

  • Tahsin Ahmed

  • Patrick Fay

  • Scott S. Howard

Journal or Work Title
  • Optics Letters

Volume
  • 42

Issue
  • 1

First Page
  • 155

Last Page
  • 158

Number of Pages
  • 3

Publication Date
  • 2016-12

Publisher
  • OSA: The Optical Society

Date Created
  • 2018-10-31

Language
  • English

Departments and Units
Record Visibility and Access Public
Content License
  • All rights reserved

Digital Object Identifier

doi:10.1364/OL.42.000155

This DOI is the best way to cite this article.