Line-scanning hyperspectral imaging based on structured illumination optical sectioning

Yu John Hsu, Chih Chiang Chen, Chien Hsiang Huang, Chia Hua Yeh, Li Ying Liu, Szu Yu Chen

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


Line-scanning hyperspectral imaging (LHSI) is known to have a higher acquisition rate but lower sectioning capability than point-scanning hyperspectral imaging. To further increase the axial imaging contrast of LHSI, structured illumination was integrated into line excitation to remove the off-focus and scattered on-focus fluorescence signals. In an unsectioned leaf, the imaging contrast can be enhanced by 8 times, while in sectioned mouse skin tissues, a 4.5-fold enhancement can be achieved. With a spectral resolution of 1.15 nm, the fluorophores with seriously-overlapped spectra was proved to be separated without cross-talk by applying linear unmixing to the recorded spectral information.

Original languageEnglish
Article number#282654
JournalBiomedical Optics Express
Issue number6
StatePublished - 1 Jun 2017


  • Fluorescence microscopy
  • Multispectral and hyperspectral imaging
  • Scanning microscopy


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