Expand this Topic clickable element to expand a topic
Skip to content
Optica Publishing Group

Saturation-compensated measurements for fluorescence lifetime imaging microscopy

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.

© 2016 Optical Society of America

Full Article  |  PDF Article
More Like This
Super-sensitivity multiphoton frequency-domain fluorescence lifetime imaging microscopy

Yide Zhang, Aamir A. Khan, Genevieve D. Vigil, and Scott S. Howard
Opt. Express 24(18) 20862-20867 (2016)

Artificial neural network approaches for fluorescence lifetime imaging techniques

Gang Wu, Thomas Nowotny, Yongliang Zhang, Hong-Qi Yu, and David Day-Uei Li
Opt. Lett. 41(11) 2561-2564 (2016)

Functional in vivo imaging using fluorescence lifetime light-sheet microscopy

Claire A. Mitchell, Simon P. Poland, James Seyforth, Jakub Nedbal, Thomas Gelot, Tahiyat Huq, Gerhard Holst, Robert D. Knight, and Simon M. Ameer-Beg
Opt. Lett. 42(7) 1269-1272 (2017)

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Figures (5)

You do not have subscription access to this journal. Figure files are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Equations (12)

You do not have subscription access to this journal. Equations are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Select as filters


Select Topics Cancel
© Copyright 2024 | Optica Publishing Group. All Rights Reserved