Three-dimensional fluorescence imaging using the transport of intensity equation

Sudheesh K. Rajput, Manoj Kumar, Xiangyu Quan, Mitsuhiro Morita, Tomoyuki Furuyashiki, Yasuhiro Awatsuji, Enrique Tajahuerce, Osamu Matoba

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

We propose a nonscanning three-dimensional (3-D) fluorescence imaging technique using the transport of intensity equation (TIE) and free-space Fresnel propagation. In this imaging technique, a phase distribution corresponding to defocused fluorescence images with a point-light-source-like shape is retrieved by a TIE-based phase retrieval algorithm. From the obtained phase distribution, and its corresponding amplitude distribution, of the defocused fluorescence image, various images at different distances can be reconstructed at the desired plane after Fresnel propagation of the complex wave function. Through the proposed imaging approach, the 3-D fluorescence imaging can be performed in multiple planes. The fluorescence intensity images are captured with the help of an electrically tunable lens; hence, the imaging technique is free from motion artifacts. We present experimental results corresponding to microbeads and a biological sample to demonstrate the proposed 3-D fluorescence imaging technique.

Original languageEnglish
Article number032004
JournalJournal of Biomedical Optics
Volume25
Issue number3
DOIs
StatePublished - 1 Mar 2020
Externally publishedYes

Keywords

  • 3-D fluorescence imaging
  • Fresnel propagation
  • phase retrieval
  • transport of intensity equation

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