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. 2017 Sep 22;8(10):4629-4639.
doi: 10.1364/BOE.8.004629. eCollection 2017 Oct 1.

Comparison and combination of rotational imaging optical coherence tomography and selective plane illumination microscopy for embryonic study

Chen Wu  1 , Henry Le  2 , Shihao Ran  3 , Manmohan Singh  1 , Irina V Larina  2 , David Mayerich  3 , Mary E Dickinson  2   4   5 , Kirill V Larin  1   2   6   4
Affiliations

Comparison and combination of rotational imaging optical coherence tomography and selective plane illumination microscopy for embryonic study

Chen Wu et al. Biomed Opt Express. .

Abstract

Several optical imaging techniques have been applied for high-resolution embryonic imaging using different contrast mechanisms, each with their own benefits and limitations. In this study, we imaged the same E9.5 mouse embryo with rotational imaging optical coherence tomography (RI-OCT) and selective plane illumination microscopy (SPIM). RI-OCT overcomes optical penetration limits of traditional OCT imaging that prohibit full-body imaging of mouse embryos at later stages by imaging the samples from multiple angles. SPIM enables high-resolution, 3D imaging with less phototoxicity and photobleaching than laser scanning confocal microscopy (LSCM) by illuminating the sample with a focused sheet of light. Side by side comparisons are supplemented with co-registered images. The results demonstrate that SPIM and RI-OCT are highly complementary and could provide more comprehensive tissue characterization for mouse embryonic research.

Keywords: (110.4500) Optical coherence tomography; (170.2520) Fluorescence microscopy.

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Figures

Fig. 1
Fig. 1
Schematic of (a) RI-OCT experimental setup and (b) SPIM setup.
Fig. 2
Fig. 2
(a) 3D OCT and (b) SPIM imaging of beads phantom. (c) Cross-sectional OCT image of selected plane. (d) Combined OCT and SPIM images of the same cross-sectional plane. Scale bars are 500 µm.
Fig. 3
Fig. 3
RI-OCT imaging of E9.5 mouse embryo at (a) 0 degree, (b) 180 degree, (c) 90 degree, (d) 270 degree, (e-h) composite from all four angles. SPIM imaging of same embryo from (i) one side, (j) opposite side, and (k-l) combined from both sides. Scale bars are 500 µm.
Fig. 4
Fig. 4
E9.5 mouse embryo. (a) 3D OCT image. (b) 3D SPIM image at an excitation wavelength of 488nm corresponding to the ϵ-globin-GFP cells (erythroblast). (c) 3D SPIM image at an excitation wavelength of 561nm for PECAM1 labeled endothelial cells. (d) 3D registered OCT and SPIM image (see Visualization 1). (e-h) 2D OCT image registered with SPIM image for various planes, and arrowhead points to the heart wall. DA: dorsal aorta, HRT: heart, EPC: ectoplacental cone, NL: neuro lumen, NVN: neuro vascular network, ISV: intersomitic vessel.
See this image and copyright information in PMC

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