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Tytuł:
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Multiphoton microscopy for label-free multicolor imaging of peripheral nerve.
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Autorzy:
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Rishøj L; Boston University, Department of Electrical and Computer Engineering, Boston, Massachusetts, United States.; Technical University of Denmark, DTU Fotonik, Kgs. Lyngby, Denmark.
Hernández IC; Mass Eye and Ear and Harvard Medical School, Surgical Photonics and Engineering Laboratory, Boston, United States.
Ramachandran S; Boston University, Department of Electrical and Computer Engineering, Boston, Massachusetts, United States.
Jowett N; Mass Eye and Ear and Harvard Medical School, Surgical Photonics and Engineering Laboratory, Boston, United States.
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Źródło:
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Journal of biomedical optics [J Biomed Opt] 2022 May; Vol. 27 (5).
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, N.I.H., Extramural
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Język:
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English
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Imprint Name(s):
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Original Publication: Bellingham, WA : Published by SPIE--the International Society for Optical Engineering in cooperation with International Biomedical Optics Society, c1996-
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MeSH Terms:
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Microscopy*
Myelin Sheath*
Animals ; Collagen ; Coloring Agents ; Mammals ; Mice ; Mice, Transgenic ; Microscopy, Fluorescence, Multiphoton/methods ; Sciatic Nerve/diagnostic imaging
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References:
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Grant Information:
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R01 NS071067 United States NS NINDS NIH HHS; R21 EY026410 United States EY NEI NIH HHS
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Contributed Indexing:
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Keywords: imaging; label-free; lasers; myelinated; nerve fibers; nonlinear optical microscopy
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Substance Nomenclature:
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0 (Coloring Agents)
9007-34-5 (Collagen)
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Entry Date(s):
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Date Created: 20220515 Date Completed: 20220517 Latest Revision: 20221012
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Update Code:
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20240105
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PubMed Central ID:
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PMC9109936
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DOI:
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10.1117/1.JBO.27.5.056501
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PMID:
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35568795
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Significance: Means for quantitation of myelinated fibers in peripheral nerve may guide diagnosis and clinical decision making in management of peripheral nerve disorders. Multiphoton microscopy techniques such as the third-harmonic generation enable label-free in vivo imaging of peripheral nerves.
Aim: Develop a multiphoton microscope based on a custom high-power infrared fiber laser for label-free imaging of peripheral nerve.
Approach: A cost-effective multiphoton microscope employing a single fiber laser source at 1300 nm was designed and used for stain-free multicolor imaging of murine and human peripheral nerve.
Results: Second-harmonic generation signal from collagen centered about 650-nm delineated neural connective tissue, whereas third-harmonic general signal centered about 433-nm delineated myelin and other lipids. In sciatic nerve from transgenic reporter mice expressing yellow fluorescent protein within peripheral neurons, three-photon-excitation with emission peak at 527-nm delineated axoplasm. The signal obtained from unlabeled axially sectioned samples was adequate for segmentation of myelinated fibers using commercial image processing software. In unlabeled whole mount specimens, imaging depths over 100-μm were achieved.
Conclusions: A multiphoton microscope powered by a fiber laser enables stain-free histomorphometry of mammalian peripheral nerve. The simplicity of the microscope design carries potential for clinical translation to inform decision making in peripheral nerve disorders.