Fast and improved bioimaging via temporal focusing multiphoton excitation microscopy with binary digital-micromirror-device holography

Yong Da Sie, Chia Yuan Chang, Chun Yu Lin, Nan Shan Chang, Paul J. Campagnola, Shean Jen Chen*

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Conventional temporal focusing-based multiphoton excitation microscopy (TFMPEM) can offer widefield optical sectioning with an axial excitation confinement of a few microns. To improve the axial confinement of TFMPEM, a binary computer-generated Fourier hologram (CGFH) via a digital-micromirror-device (DMD) was implemented to intrinsically improve the axial confinement by filling the back-focal aperture of the objective lens. Experimental results show that the excitation focal volume can be condensed and the axial confinement improved about 24% according to the DMD holography. In addition, pseudouniform MPE can be achieved using two complementary CGFHs with rapid pulse-width modulation switching via the DMD. Furthermore, bioimaging of CV-1 in origin with SV40 genes-7 cells demonstrates that the TFMPEM with binary DMD holography can improve image quality by enhancing axial excitation confinement and rejecting out-of-focus excitation.

Original languageEnglish
Article number116502
JournalJournal of Biomedical Optics
Volume23
Issue number11
DOIs
StatePublished - 1 Jan 2018

Keywords

  • fluorescence microscopy
  • medical and biological imaging
  • nonlinear microscopy
  • ultrafast lasers

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