Details
| Original language | English |
|---|---|
| Title of host publication | Three-Dimensional and Multidimensional Microscopy |
| Subtitle of host publication | Image Acquisition and Processing XXV |
| Publisher | SPIE |
| ISBN (electronic) | 9781510614833 |
| Publication status | Published - 23 Feb 2018 |
| Event | Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV 2018 - San Francisco, United States Duration: 29 Jan 2018 → 31 Jan 2018 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 10499 |
| ISSN (Print) | 1605-7422 |
Abstract
Scanning laser optical tomography (SLOT) is a 3D imaging technique, based on the principle of computed tomography to visualize samples up to magnitude of several centimeters. Intrinsic contrast mechanisms as absorption, scattering and autofluorescence provide information about the 3D architecture and composition of the sample. Another valuable intrinsic contrast mechanism is second harmonic generation (SHG), which is generated in noncentrosymmetric materials and commonly used to image collagen in biological samples. The angular dependence of the SHG signal, however, produces artifacts in reconstructed optical tomography datasets (OPT, SLOT). Thus, successful use of this intrinsic contrast mechanism is impaired. We investigate these artifacts by simulation and experiment and propose an elimination procedure that enables successful reconstruction of SHG-SLOT data. Nevertheless, in many cases specific labeling of certain structures is necessary to make them visible. Using multiple dyes in one sample can lead to crosstalk between the different channels and reduce contrast of the images. Also autofluorescence of the sample itself can account for that. By using multispectral imaging in combination with spectral unmixing techniques, this loss can be compensated. Therefore either a spectrally resolved detection path, or spectrally resolved excitation is required. Therefore we integrated a white supercontinuum light source in our SLOT-setup that enables a spectral selection of the excitation beam and extended the detection path to a four channel setup. This enables the detection of three fluorescence channels and one absorption channel in parallel, and increases the contrast in the reconstructed 3D images significantly.
Keywords
- 3D Microscopy, Multispectral imaging, Nonlinear Optics, Scanning Laser Optical Tomography, Second Harmonic Generation, Spectral unmixing, Tomographic imaging
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Materials Science(all)
- Biomaterials
- Physics and Astronomy(all)
- Atomic and Molecular Physics, and Optics
- Medicine(all)
- Radiology Nuclear Medicine and imaging
Cite this
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- Harvard
- Apa
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- BibTeX
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Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV. SPIE, 2018. 1049913 (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 10499).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Investigation of second harmonic generation and multispectral imaging as new contrast mechanisms in scanning laser optical tomography
AU - Nolte, Lena
AU - Antonopoulos, Georgios C.
AU - Heisterkamp, Alexander
AU - Ripken, Tammo
AU - Meyer, Heiko
N1 - Funding information: This work was supported by the German Federal Ministry of Education and Research (Pionieraxon: 031L0062B) and by the Ministry of Lower Saxony and the VolkswagenStiftung (both BIOFABRICATION FOR NIFE: VWZN2860, http://biofabrication.info/).
PY - 2018/2/23
Y1 - 2018/2/23
N2 - Scanning laser optical tomography (SLOT) is a 3D imaging technique, based on the principle of computed tomography to visualize samples up to magnitude of several centimeters. Intrinsic contrast mechanisms as absorption, scattering and autofluorescence provide information about the 3D architecture and composition of the sample. Another valuable intrinsic contrast mechanism is second harmonic generation (SHG), which is generated in noncentrosymmetric materials and commonly used to image collagen in biological samples. The angular dependence of the SHG signal, however, produces artifacts in reconstructed optical tomography datasets (OPT, SLOT). Thus, successful use of this intrinsic contrast mechanism is impaired. We investigate these artifacts by simulation and experiment and propose an elimination procedure that enables successful reconstruction of SHG-SLOT data. Nevertheless, in many cases specific labeling of certain structures is necessary to make them visible. Using multiple dyes in one sample can lead to crosstalk between the different channels and reduce contrast of the images. Also autofluorescence of the sample itself can account for that. By using multispectral imaging in combination with spectral unmixing techniques, this loss can be compensated. Therefore either a spectrally resolved detection path, or spectrally resolved excitation is required. Therefore we integrated a white supercontinuum light source in our SLOT-setup that enables a spectral selection of the excitation beam and extended the detection path to a four channel setup. This enables the detection of three fluorescence channels and one absorption channel in parallel, and increases the contrast in the reconstructed 3D images significantly.
AB - Scanning laser optical tomography (SLOT) is a 3D imaging technique, based on the principle of computed tomography to visualize samples up to magnitude of several centimeters. Intrinsic contrast mechanisms as absorption, scattering and autofluorescence provide information about the 3D architecture and composition of the sample. Another valuable intrinsic contrast mechanism is second harmonic generation (SHG), which is generated in noncentrosymmetric materials and commonly used to image collagen in biological samples. The angular dependence of the SHG signal, however, produces artifacts in reconstructed optical tomography datasets (OPT, SLOT). Thus, successful use of this intrinsic contrast mechanism is impaired. We investigate these artifacts by simulation and experiment and propose an elimination procedure that enables successful reconstruction of SHG-SLOT data. Nevertheless, in many cases specific labeling of certain structures is necessary to make them visible. Using multiple dyes in one sample can lead to crosstalk between the different channels and reduce contrast of the images. Also autofluorescence of the sample itself can account for that. By using multispectral imaging in combination with spectral unmixing techniques, this loss can be compensated. Therefore either a spectrally resolved detection path, or spectrally resolved excitation is required. Therefore we integrated a white supercontinuum light source in our SLOT-setup that enables a spectral selection of the excitation beam and extended the detection path to a four channel setup. This enables the detection of three fluorescence channels and one absorption channel in parallel, and increases the contrast in the reconstructed 3D images significantly.
KW - 3D Microscopy
KW - Multispectral imaging
KW - Nonlinear Optics
KW - Scanning Laser Optical Tomography
KW - Second Harmonic Generation
KW - Spectral unmixing
KW - Tomographic imaging
UR - http://www.scopus.com/inward/record.url?scp=85045318381&partnerID=8YFLogxK
U2 - 10.1117/12.2289509
DO - 10.1117/12.2289509
M3 - Conference contribution
AN - SCOPUS:85045318381
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Three-Dimensional and Multidimensional Microscopy
PB - SPIE
T2 - Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV 2018
Y2 - 29 January 2018 through 31 January 2018
ER -