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Published on: October 23, 2017
Single myelin fiber imaging in living rodents without labeling by deep optical coherence microscopy
Juliette Ben Arous1, Jonas Binding, Jean-François Léger
1Ecole Normale Supérieure, Institut de Biologie de l'ENS, IBENS 46 rue d'Ulm, Paris 75005 [corrected] France.
Abstract:
Myelin sheath disruption is responsible for multiple neuropathies in the central and peripheral nervous system. Myelin imaging has thus become an important diagnosis tool. However, in vivo imaging has been limited to either low-resolution techniques unable to resolve individual fibers or to low-penetration imaging of single fibers, which cannot provide quantitative information about large volumes of tissue, as required for diagnostic purposes. Here, we perform myelin imaging without labeling and at micron-scale resolution with >300-μm penetration depth on living rodents. This was achieved with a prototype [termed deep optical coherence microscopy (deep-OCM)] of a high-numerical aperture infrared full-field optical coherence microscope, which includes aberration correction for the compensation of refractive index mismatch and high-frame-rate interferometric measurements. We were able to measure the density of individual myelinated fibers in the rat cortex over a large volume of gray matter. In the peripheral nervous system, deep-OCM allows, after minor surgery, in situ imaging of single myelinated fibers over a large fraction of the sciatic nerve. This allows quantitative comparison of normal and Krox20 mutant mice, in which myelination in the peripheral nervous system is impaired. This opens promising perspectives for myelin chronic imaging in demyelinating diseases and for minimally invasive medical diagnosis.
Insights
Deep optical coherence microscopy enables high-resolution myelin imaging in living rodents without labels. This breakthrough allows detailed visualization of nerve fibers for diagnosing neuropathies and demyelinating diseases.
Area of Science:
- Neuroscience
- Biomedical Imaging
- Optical Microscopy
Background:
- Myelin sheath disruption causes various neuropathies.
- Current in vivo myelin imaging techniques lack resolution, penetration depth, or quantitative capabilities for diagnostics.
- Accurate myelin imaging is crucial for diagnosing central and peripheral nervous system disorders.
Purpose of the Study:
- To develop a novel imaging technique for high-resolution, deep-penetration myelin imaging in living animals.
- To overcome limitations of existing in vivo myelin imaging methods.
- To enable quantitative assessment of myelinated fibers in both the central and peripheral nervous systems.
Main Methods:
- Utilized a prototype deep optical coherence microscopy (deep-OCM) system.
- Employed a high-numerical aperture infrared full-field optical coherence microscope with aberration correction.
- Performed high-frame-rate interferometric measurements for label-free myelin imaging.
Main Results:
- Achieved micron-scale resolution myelin imaging with >300-μm penetration depth in living rodents.
- Successfully measured the density of individual myelinated fibers in rat cortex gray matter over large volumes.
- Enabled in situ imaging of single myelinated fibers in the sciatic nerve of Krox20 mutant mice, revealing myelination defects.
Conclusions:
- Deep-OCM provides unprecedented capabilities for label-free, high-resolution myelin imaging in vivo.
- This technique facilitates quantitative analysis of myelinated nerve fibers in both the central and peripheral nervous systems.
- Deep-OCM holds significant promise for chronic imaging in demyelinating diseases and minimally invasive medical diagnosis.
