A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

Simon P. Poland, Grace Chan, James Levitt, Nikola Krstajić, Ahmet Erdogan, Robert Henderson, Maddy Parsons, Simon Ameer-Beg

Research output: Contribution to journalArticlepeer-review

Abstract

In this paper we will discuss the development of a
multifocal multiphoton fluorescent lifetime imaging
system where 4 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
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 pre-programmed
by the user. We demonstrate the capabilities of the
system to image live-cell interactions. Whilst only 4
planes are shown, this technique can be rescaled to a
large number of focal planes enabling full 3D acquisition
and reconstruction.
Original languageEnglish
Pages (from-to)6057-6060
Number of pages4
JournalOptics Letters
Volume43
Issue number24
Early online date5 Nov 2018
DOIs
Publication statusPublished - 15 Dec 2018

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