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Sci Rep. 2017 Aug 17;7(1):8591. doi: 10.1038/s41598-017-08822-0.

Oblique scanning laser microscopy for simultaneously volumetric structural and molecular imaging using only one raster scan.

Author information

1
Department of Medicine, Boston University School of Medicine, Boston, MA, 02118, USA.
2
Center of Regenerative Medicine, Boston University, Boston, MA, 02118, USA.
3
Department of Medicine, Boston University School of Medicine, Boston, MA, 02118, USA. jiyi@bu.edu.
4
Center of Regenerative Medicine, Boston University, Boston, MA, 02118, USA. jiyi@bu.edu.
5
Boston University Photonics Center, Boston, MA, 02215, USA. jiyi@bu.edu.

Abstract

Multi-modal three dimensional (3D) optical imaging combining both structural sensitivity and molecular specificity is highly desirable in biomedical research. In this paper, we present a method termed oblique scanning laser microscopy (OSLM) to combine optical coherence tomography (OCT), for simultaneously volumetric structural and molecular imaging with cellular resolution in all three dimensions. Conventional 3D laser scanning fluorescence microscopy requires repeated optical sectioning to create z-stacks in depth. Here, the use of an obliquely scanning laser eliminates the z-stacking process, then allows highly efficient 3D OCT and fluorescence imaging by using only one raster scan. The current setup provides ~3.6 × 4.2 × 6.5 μm resolution in fluorescence imaging, ~7 × 7 × 3.5 μm in OCT in three dimensions, and the current speed of imaging is up to 100 frames per second (fps) over a volume about 0.8 × 1 × 0.5 mm3. We demonstrate several mechanisms for molecular imaging, including intrinsically expressed GFP fluorescence, autofluorescence from Flavin proteins, and exogenous antibody-conjugated dyes. We also demonstrate potential applications in imaging human intestinal organoids (HIOs), colon mucosa, and retina.

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