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Multifocal multiphoton volumetric imaging approach for high-speed time-resolved Förster resonance energy transfer imaging in vivo

Authors
  • Poland, Simon P.
  • Chan, Grace K.
  • Levitt, James A.
  • Krstajić, Nikola
  • Erdogan, Ahmet T.
  • Henderson, Robert K.
  • Parsons, Maddy
  • Ameer-Beg, Simon M.
Type
Published Article
Journal
Optics Letters
Publisher
The Optical Society
Publication Date
Dec 12, 2018
Volume
43
Issue
24
Pages
6057–6060
Identifiers
DOI: 10.1364/OL.43.006057
PMID: 30548010
PMCID: PMC6410918
Source
PubMed Central
License
Unknown

Abstract

In this Letter, we will discuss the development of a multifocal multiphoton fluorescent lifetime imaging system where four individual fluorescent intensity and lifetime planes are acquired simultaneously, allowing us to obtain volumetric data without the need for sequential scanning at different axial depths. Using a phase-only spatial light modulator (SLM) with an appropriate algorithm to generate a holographic pattern, we project a beamlet array within a sample volume of a size, which can be preprogrammed by the user. We demonstrate the capabilities of the system to image live-cell interactions. While only four planes are shown, this technique can be rescaled to a large number of focal planes, enabling full 3D acquisition and reconstruction.

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