Combined optical coherence tomography and fluorescent two-photon selective-plane illumination microscopy for embryonic imaging
Md Mobarak Karim1, Ruijiao Sun2, Behzad Khajavi1, Manmohan Singh1, Harshdeep S. Chawla1, Alexander W. Schill1, David Mayerich2, Mary E. Dickinson3, and Kirill V Larin1,3,*
1 Department of Biomedical Engineering, University of Houston, Houston, TX, USA
2 Department of Electrical and Computer Engineering, University of Houston, Houston, TX, USA
3 Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA
* klarin@uh.edu
Abstract
We demonstrate the combination of Optical Coherence Tomography (OCT) and Fluorescent Two-Photon Selective-Plane Illumination Microscopy (2P-SPIM) for high-resolution multimodal embryonic imaging. 2P-SPIM enables deeper penetration and lower phototoxicity and photobleaching than laser scanning confocal microscopy, enabling more robust longitudinal imaging. The 2P-SPIM collection arm consists of a 16X water immersion objective, tube lens, and CCD camera, resulting in a resolution of ~2.1 µm. OCT imaging provides label-free micrometer-resolution structural imaging along the same plane as the 2P-SPIM system. The OCT system is based on a 100 kHz swept-source laser with a central wavelength of 1050 nm and has a lateral resolution of ~15 µm and an axial resolution of ~7 µm. The unique combination of OCT and 2P-SPIM enables deep imaging of embryo structure and function, such as cell proliferation, cell type identification, and cell signaling. First, we imaged green fluorescence microspheres (1-5 µm) and red fluorescence microspheres (1-5 µm) embedded in phantoms to demonstrate the capabilities of the multimodal system. Next, we imaged a mouse embryo with the combined OCT and 2P-SPIM system to demonstrate its multimodal imaging capabilities in tissues. With OCT and 2P-SPIM, we have the capability for easily co-registered structural and functional imaging in a single instrument.
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Md Mobarak Karim
Department of Biomedical Engineering, University of Houston, Houston, TX
USA
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