Multimodal optical coherence tomography and selective-plane illumination microscopy for embryonic imaging

Md Mobarak Karim
SEC 206
Fluorescent selective-plane illumination microscopy (SPIM) can enable to provide cell information
such as cell proliferation, cell type identification, and cell signaling while optical coherence tomography
(OCT) can capture the complementary structural information. We developed very first multimodal highresolution
embryonic imaging system to combine the benefits of OCT with SPIM. SPIM allows highresolution
3D imaging with low phototoxicity and photobleaching than laser scanning confocal
microscopy which enhances more robust information from the targeted sample. The OCT and SPIM
beams were coupled by using a beam splitter. Utilizing a galvanometer-mounted mirror and the same
objective lens, the OCT probe and LSFM excitation beams were merged and scanned. The resulting
light sheet thickness was ~13 μm and transverse resolution of SPIM was ~2.1 μm. With a lateral
resolution of 15 μm and an axial resolution of 7 μm, the Michelson interferometer-based swept source
OCT system provides structural information on the same plane as SPIM. The capabilities of the
multimodal imaging system were demonstrated using images of fluorescent microbeads and a
fluorescently tagged mouse embryo at gestational day 9.5. Due to the co-alignment of the OCT and
SPIM systems, image registration was simple and allowed for high-throughput multimodal imaging
without the use of sophisticated registration methods.