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

Simon P. Poland, Grace K. Chan, James A. Levitt, Nikola Krstajic, Ahmet T. Erdogan, Robert K. Henderson, Maddy Parsons, Simon M. Ameer-Beg

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    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.
    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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    Cite this

    Poland, S. P., Chan, G. K., Levitt, J. A., Krstajic, N., Erdogan, A. T., Henderson, R. K., Parsons, M., & Ameer-Beg, S. M. (2018). Multifocal multiphoton volumetric imaging approach for high-speed time-resolved Förster resonance energy transfer imaging in vivo. Optics Letters, 43(24), 6057-6060. https://doi.org/10.1364/OL.43.006057