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

Single pulse two-photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate and an all fiber based setup

Not Accessible

Your library or personal account may give you access

Abstract

Newly developed microscopy methods have the goal to give researches in bio-molecular science a better understanding of processes ongoing on a cellular level. Especially two-photon excited fluorescence (TPEF) microscopy is a readily applied and widespread modality. Compared to one photon fluorescence imaging, it is possible to image not only the surface but also deeper lying structures. Together with fluorescence lifetime imaging (FLIM), which provides information on the chemical composition of a specimen, deeper insights on a molecular level can be gained. However, the need for elaborate light sources for TPEF and speed limitations for FLIM hinder an even wider application. In this contribution, we present a way to overcome this limitations by combining a robust and inexpensive fiber laser for nonlinear excitation with a fast analog digitization method for rapid FLIM imaging. The applied sub nanosecond pulsed laser source is perfectly suited for fiber delivery as typically limiting non-linear effects like self-phase or cross-phase modulation (SPM, XPM) are negligible. Furthermore, compared to the typically applied femtosecond pulses, our longer pulses produce much more fluorescence photons per single shot. In this paper, we show that this higher number of fluorescence photons per pulse combined with a high analog bandwidth detection makes it possible to not only use a single pulse per pixel for TPEF imaging but also to resolve the exponential time decay for FLIM. To evaluate our system, we acquired FLIM images of a dye solution with single exponential behavior to assess the accuracy of our lifetime determination and also FLIM images of a plant stem at a pixel rate of 1 MHz to show the speed performance of our single pulse two-photon FLIM (SP-FLIM) system.

© 2017 SPIE

PDF Article
More Like This
High-Speed Two-Photon Fluorescence Lifetime Imaging Microscopy of NADH for Label-Free Metabolic Imaging

Andrew J. Bower, Joanne Li, Eric J. Chaney, Marina Marjanovic, and Stephen A. Boppart
OmS2D.5 Optical Molecular Probes, Imaging and Drug Delivery (OMP) 2017

Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

Rakesh Patalay, Clifford Talbot, Yuriy Alexandrov, Ian Munro, Hans Georg Breunig, Karsten König, Sean Warren, Mark A. A. Neil, Paul M. W. French, Anthony Chu, Gordon W. Stamp, and Chris Dunsby
808718 European Conference on Biomedical Optics (ECBO) 2011

Select as filters


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