Enabling second harmonic generation as a contrast mechanism for optical projection tomography (OPT) and scanning laser optical tomography (SLOT)
| Autor: | Alexander Heisterkamp, Georgios C. Antonopoulos, Tammo Ripken, Lena Nolte, Lisa Rämisch, Heiko Meyer |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Materials science Microscope animal structures Nonlinear optics Reconstruction artifacts Laser scanning Biomedical signal processing Superconducting materials Light scattering Fluorescence law.invention 03 medical and health sciences Optics Non-centrosymmetric law Microscopy medicine Second harmonic generation microscopies (SHG) High harmonic generation ddc:530 Scanning High resolution image Optical tomography Musculoskeletal system Tissue medicine.diagnostic_test Harmonic generation business.industry Second-harmonic generation Laser Computerized tomography Atomic and Molecular Physics and Optics 030104 developmental biology Scattering and absorption Isotropic resolution Simultaneous acquisition Collagen Dewey Decimal Classification::500 | Naturwissenschaften::530 | Physik business Optical projection tomography Biotechnology |
| Zdroj: | Biomedical Optics Express 9 (2018), Nr. 6 |
| ISSN: | 2156-7085 |
| Popis: | Volumetric imaging of connective tissue provides insights into the structure of biological tissue. Second harmonic generation (SHG) microscopy has become a standard method to image collagen rich tissue like skin or cornea. Due to the non-centrosymmetric architecture, no additional label is needed and tissue can be visualized noninvasively. Thus, SHG microscopy enables the investigation of collagen associated diseases, providing high resolution images and a field of view of several hundreds of µm. However, the in toto visualization of larger samples is limited to the working distance of the objective and the integration time of the microscope setup, which can sum up to several hours and days. A faster imaging technique for samples in the mesoscopic range is scanning laser optical tomography (SLOT), which provides linear fluorescence, scattering and absorption as intrinsic contrast mechanisms. Due to the advantages of SHG and the reduced measurement time of SLOT, the integration of SHG in SLOT would be a great extension. This way SHG measurements could be performed faster on large samples, providing isotropic resolution and simultaneous acquisition of all other contrast mechanisms available, such as fluorescence and absorption. SLOT is based on the principle of computed tomography, which requires the rotation of the sample. The SHG signal, however, depends strongly on the sample orientation and the polarization of the laser, which results in SHG intensity fluctuation during sample rotation and prevents successful 3D reconstruction. In this paper we investigate the angular dependence of the SHG signal by simulation and experiment and found a way to eliminate reconstruction artifacts caused by this angular dependence in SHG-SLOT data. This way, it is now possible to visualize samples in the mesoscopic range using SHG-SLOT, with isotropic resolution and in correlation to other contrast mechanisms as absorption, fluorescence and scattering. |
| Databáze: | OpenAIRE |
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