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Swept source optical coherence microscopy using a Fourier domain mode-locked laser

Shu-Wei Huang, Aaron D. Aguirre, Robert A. Huber, Desmond C. Adler, and James G. Fujimoto

Open Access Open Access

Abstract

Swept source optical coherence microscopy (OCM) enables cellular resolution en face imaging as well as integration with optical coherence tomography (OCT) cross sectional imaging. A buffered Fourier domain mode-locked (FDML) laser light source provides high speed, three dimensional imaging. Image resolutions of 1.6 μm × 8 μm (transverse × axial) with a 220 μm × 220 μm field of view and sensitivity higher than 98 dB are achieved. Three dimensional cellular imaging is demonstrated in vivo in the Xenopus laevis tadpole and ex vivo in the rat kidney and human colon.

©2007 Optical Society of America

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Figures (6)
Fig. 1.
Fig. 1. Schematic of the swept source OCM system. L1-L4, lenses; M1, mirror; OB, water-immersion objective; D1-D2, dual-balanced photodetectors; FW, neutral density filter wheel; DC, dispersion compensating glass.

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Fig. 2.
Fig. 2. System characterization. A. The confocal gate (solid line) and the coherence gate (dashed line) are measured to be ∼20 μm and ∼8 μm, respectively. B. Transverse resolution is estimated to be ∼1.6 μm and the smallest elements on the USAF resolution chart can be resolved. C. Measured sensitivity is higher than 98 dB and dynamic range is higher than 50 dB.

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Fig. 3.
Fig. 3. Extraction of en face images from the 3D dataset. A. A volumetric image acquired by the swept source OCM system in ∼1.5 second. A series of en face images can then be extracted from the volume digitally. The volume size is ∼220 μm × 220 μm × 220 μm. B, C, and D. Three representative en face images ∼12 μm apart in depth from each other are extracted from the volume around the focus.

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Fig. 4.
Fig. 4. In vivo cellular images of a Xenopus laevis tadpole. A, C. OCM images at ∼200 μm and ∼400 μm below the surface, respectively. B, D. Confocal-like images generated by summing the 3D datasets in the axial direction. Obscuration of detailed cellular structures and loss of contrast in the confocal-like images are evident. Scale bar: 50 μm.

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Fig. 5.
Fig. 5. Cellular images of a fixed rat kidney. A, B. OCM images at ∼40 μm and ∼120 μm below the surface, respectively. C. Representative histology stained with H&E. The cell lining along the kidney tubules and nuclei can be observed. Scale bar: 50 μm.

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Fig. 6.
Fig. 6. Cellular images of an unfixed human colonic mucosa. A. OCM image of a single crypt structure at ∼ 100 μm below the surface. B. Representative histology stained with H&E. C, D. OCM images of a different region at ∼100 μm and ∼150 μm below the surface, respectively. Features like round crypts, goblect cells, epithelium lining the lumen, lymphoid cells in the lamina propria, and lumen shrinkage over depth can be resolved. Scale bar: 50 μm.

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