The DOI matches its Crossref record; 65 of the 66 references it cites were found in a registry or an index; 12 of the 36 sources its claims cite were checked for support.
The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains
Open accessAbstract
Diffusion tensor imaging (DTI) is used extensively in neuroscience to noninvasively estimate white matter (WM) microarchitecture. However, the diffusion signal is inherently ambiguous because it infers WM structure from the orientation of water diffusion and cannot identify the biological sources of diffusion changes. To compare inferred WM estimates to directly labeled axonal elements, we performed a novel within-subjects combination of high-resolution ex vivo DTI with two-photon laser microscopy of intact mouse brains rendered optically transparent by Clear Lipid-exchanged, Anatomically Rigid, Imaging/immunostaining compatible, Tissue hYdrogel (CLARITY). We found that myelin basic protein (MBP) immunofluorescence significantly correlated with fractional anisotropy (FA), especially in WM regions with coherent fiber orientations and low fiber dispersion. Our results provide evidence that FA is particularly sensitive to myelination in WM regions with these characteristics. Furthermore, we found that radial diffusivity (RD) was only sensitive to myelination in a subset of WM tracts, suggesting that the association of RD with myelin should be used cautiously. This combined DTI-CLARITY approach illustrates, for the first time, a framework for using brain-wide immunolabeling of WM targets to elucidate the relationship between the diffusion signal and its biological underpinnings. This study also demonstrates the feasibility of a within-subject combination of noninvasive neuroimaging and tissue clearing techniques that has broader implications for neuroscience research.
Overview
AI snapshot
Written from the paper’s full text.
TL;DR
Combining ex vivo DTI with CLARITY whole-brain myelin basic protein (MBP) immunofluorescence in the same mouse brains, the authors found that FA correlates with MBP, especially in coherent, highly myelinated white matter regions. RD correlated with MBP only in a subset of tracts, indicating that RD–myelin associations should be interpreted cautiously.
Key findings
- CLARITY MBP-labeled ROI volumes showed excellent structural correspondence with DTI ROIs (Spearman’s ρ=0.876, P<0.00001).
- Across all WM ROIs, FA significantly correlated with MBP immunofluorescence (Χ²(1)=7.74, P<0.01), whereas AD (Χ²(1)=0.27, P=0.61) and RD (Χ²(1)=0.05, P=0.83) did not.
- In a WM subset including corpus callosum, fimbria, and anterior commissure, MBP positively correlated with FA (Χ²(1)=12.56, P<0.0005) and negatively with RD (Χ²(1)=10.54, P<0.005); in remaining tracts, FA still correlated with MBP (Χ²(1)=10.62, P<0.005) but RD did not (P=0.37).
- Within the stria medullaris, FA–MBP correlation was significant in high-FA ROIs >0.5 (ρ=0.666, P<0.00001, n=44 regions) but not in low-FA ROIs (P=0.118); high-FA regions had higher linear anisotropy (Cl=0.51±0.021) than diverging-fascicle regions (Cl=0.35±0.024).
Methods
- Within-subjects design combining ex vivo DTI at 11.7 T with CLARITY/PACT tissue clearing and two-photon microscopy of MBP immunofluorescence in intact mouse brains.
- Main experiments used four male C57BL/6 J mice, 3–6 months old; three male thy1-eYFP-H mice were used for initial pilot compatibility testing.
- DTI used a 3D DW-GRASE sequence, 15 directions, b=1500 s/mm², 100 μm isotropic resolution, ~16.5 h scan time; 14 discrete 3D WM ROIs were traced in DTI and CLARITY spaces, and mixed-effect models tested DTI metrics as predictors of MBP with ROI as fixed effect and subject as random effect.
Limitations
- The paper states that incomplete antibody penetration or uneven labeling may leave some myelinated areas uncaptured; fixation and the liquid hydrogel may affect diffusivity in ways that cannot be fully ruled out; and MBP immunofluorescence quantification in fixed tissue must be interpreted cautiously, with electron microscopy needed for further validation.
- The paper also notes that the FA–myelin relationship is complex and cannot be resolved by this correlational approach alone, and that MBP expression does not necessarily index myelin sheath thickness or the percentage of myelinated axons.
- Evident from the text: the main sample is small (four mice), imaging is ex vivo at 28 °C, and even at 100 μm resolution each voxel may contain 1,000–13,000 axons, over 80 oligodendrocytes, and more than 20 astrocytes, limiting biological specificity.
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Claims the paper makes
27 Own result · 12 Own interpretation · 10 No full text of the sources · 6 Partly supported · 4 Own method · 3 Supported · 1 Supported so far · 1 Couldn’t confirm · 1 Sources not found · 1 Not checked yet · 1 No basis given · accuracy not yet measured
Every source the paper cites for this claim was checked and supports it.
In the paper
“This demonstrated that CLARITY could be successfully performed despite the presence of a contrast agent in the hydrogel and minor deviations from the original published protocol (Chung et al., 2013).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.1. CLARITY polymerization and tissue clearing are normal following ex vivo DTI
What the sources say
- Supportedcites Chung et al., 2013 source
The citation identifies the original CLARITY protocol against which the citing authors describe their modifications. The source’s Methods (“Basic CLARITY protocol”; page unavailable) provides that protocol. The contrast-agent result is the citing authors’ own conclusion, not a finding attributed to Chung et al. (LLM, conf=0.86)
boostedn = 3- Asserted: passes — The authors put it forward as true.
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- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“When coupled with two-photon or light-sheet laser microscopy, whole mouse brains can be imaged entirely intact with sub-cellular resolution (Tomer et al., 2014).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
- Partly supportedcites Tomer et al., 2014 sourceExperimental — accuracy not yet measured
The Overview of procedure and Anticipated Results (page unavailable) report high-resolution imaging through an intact mouse brain and whole-brain imaging with CLARITY-optimized light-sheet microscopy. The paper does not demonstrate whole-brain imaging with two-photon microscopy or establish sub-cellular resolution throughout the whole brain. (LLM, conf=0.86)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Partly supported
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In the paper
“This has led to considerable debate, since the inception of DTI, over the biological significance of these DTI-derived measures since WM integrity can be affected by many different parameters including axon packing density, axon caliber, myelination, microglia, inflammation, and tissue architecture (Mori and Zhang, 2006; Le Bihan and Johansen-Berg, 2012; Jones et al., 2013).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
- Not checked — no full text of the sourcecites Mori and Zhang, 2006 source
- Partly supportedcites Le Bihan and Johansen-Berg, 2012 sourceExperimental — accuracy not yet measured
The section on diffusion anisotropy in white matter links the measure to fiber packing, membrane organization, myelin, and tissue orientation; the section on brain development also discusses axon density and diameter. The cited paper supports the broader point that DTI measures have multiple biological influences, but our copy does not identify microglia or inflammation as influences on white-matter integrity or these measures (searched: “microglia” and “inflammation”). No page numbers were provided for these passages. (LLM, conf=0.88)
- Not checked — no full text of the sourcecites Jones et al., 2013 source
plaindirect- Asserted: passes — The authors put it forward as true.
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Every source the paper cites for this claim was checked and supports it.
In the paper
“It has been estimated that up to 90% of WM voxels in a DTI map contain complex fiber architecture and cannot be modeled accurately using a single tensor model (Jeurissen et al., 2013).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
What the sources say
- Supportedcites Jeurissen et al., 2013 source
The Discussion reports multiple fiber orientations in approximately 90% of white-matter voxels using CSD. The Introduction states that the diffusion tensor model is inadequate in voxels containing multiple fiber orientations. The 90% estimate comes from CSD analysis of diffusion data, rather than detection by DTI itself. (LLM, conf=0.92)
hedged- Asserted: passes — The authors put it forward as true.
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- Supported
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Moving from macroscopic to microscopic scales further highlights the discrepancy between the principal eigenvectors and the physical fiber bundles that are actually present at those locations in the brain (Fig. 5A and B).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“and that MBP expression within these structures significantly correlated with DTI-derived FA measures.”
- Also stated in: § 3.3. CLARITY MBP correlates with DTI-derived FA measures · plain
“we found, using a mixed effect model, that FA significantly correlated with MBP immunofluorescence across the full set of ROIs (Fig. 3C; Χ2(1)=7.74, P<0.01).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
- Rests on an assumption: The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
- Rests on an assumption: The ex vivo DTI scanning and CLARITY hydrogel perfusion do not alter the relationship between MBP and DTI metrics.
- Also stated in: § 3.3. CLARITY MBP correlates with DTI-derived FA measures · plain
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Geometric anisotropy indices estimated that the body of the corpus collosum contained more crossing or mixing fiber architecture as reflected by its lower linear anisotropy (Cl=0.36 ± 0.030) and higher planar anisotropy (Cp=0.29 ± 0.031), compared to the genu and splenium (genu: Cl=0.53 ± 0.033, Cp=0.17 ±0.019; splenium, Cl=0.48±0.049, Cp=0.19 ±0.011).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
- Rests on an assumption: The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“we have demonstrated that FA measures significantly correlate with a direct label of axon myelination.”
- Also stated in: § 5. Conclusions · plain
“By using MBP as a proxy for myelination, we showed that it correlated with DTI-derived FA across intact WM structures, particularly in coherent and highly organized fiber regions.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
boostedcorrelational- Asserted: passes — The authors put it forward as true.
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- Based: passes — The paper gives a reason why it holds.
- Also stated in: § 5. Conclusions · plain
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“There was a highly significant positive correlation between the volume of CLARITY MBP-labeled structures and volumes for these same ROIs in DTI (Fig. 3B; Spearman's correlation, ρ=0.876, P < 0.00001)”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.3. CLARITY MBP correlates with DTI-derived FA measures
boostedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“This study also demonstrates the feasibility of a within-subject combination of noninvasive neuroimaging and tissue clearing techniques that has broader implications for neuroscience research.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > Abstract
boosted- Asserted: passes — The authors put it forward as true.
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- It may state two separate claims as one. 19
- Probably not a substantial claim. 9
Every source the paper cites for this claim was checked and supports it.
In the paper
“CLARITY achieves tissue transparency to enable deep laser penetration in normally highly light-scattering mouse brains, while preserving proteins in situ and the native 3D structure of brain tissue.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
- Supportedcites Chung et al., 2013 source
The Whole adult mouse brain imaging section reports that CLARITY makes the intact mouse brain transparent, permits deep optical imaging, and retains fine structural details without net tissue deformation; page numbers are unavailable in our copy. The Molecular phenotyping section reports only about 8% protein loss after clearing, supporting preservation of proteins in place. (LLM, conf=0.93)
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Clear Lipid-exchanged, Anatomically Rigid, Imaging/immunostaining compatible, Tissue hYdrogel (CLARITY; Chung et al., 2013) is a clearing technique that transforms intact brain tissue into nanoporous, lipid-free, hydrogel-tissue hydrids that are optically transparent but macromolecule permeable.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
- Partly supportedcites Chung et al., 2013 sourceExperimental — accuracy not yet measured
The Abstract describes CLARITY as transforming intact tissue into a nanoporous, optically transparent, macromolecule-permeable hydrogel hybrid; the ‘Hydrogel-electrophoretic tissue transmutation’ section describes lipid extraction. However, the citing sentence expands the acronym differently from the paper’s Methods > CLARITY section, which specifies ‘Acrylamide-hybridized Rigid’ rather than ‘Anatomically Rigid.’ (LLM, conf=0.84)
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“However, even at this 100 μm resolution and ∼17 h scan times, each imaging voxel can contain anywhere from 1,000 to 13,000 axons, over 80 oligodendrocytes, and more than 20 astrocytes (Walhovd et al., 2014).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Partly supportedcites Walhovd et al., 2014 sourceExperimental — accuracy not yet measured
In “Toward a multiscale understanding: From a myelin biology point of view, what does a voxel contain?” (page unavailable), the paper reports 1,000 to over 13,000 axons and averages of 86 oligodendrocytes and 22 astrocytes in a 100 μm-cubed volume of rodent white matter. These support the cited counts, though the glial figures are averages rather than counts guaranteed for each voxel. The passages found do not substantiate the ∼17-hour scan time. (LLM, conf=0.86)
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
- Partly supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“Lastly, we have used MBP immunofluorescence as a marker of myelination, however, multiple variables can influence the quantitation of fluorophores in fixed tissue samples (Straight, 2007; Waters, 2009) and this interpretation must be considered cautiously.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Not checked — no full text of the sourcecites Straight, 2007 source
- Supportedcites Waters, 2009 source
The Specimen preparation section cautions that fixation can alter detectable epitopes and that antibody penetration may vary across tissue, so fluorescence intensity may not reflect epitope quantity. This supports the cited caution about quantifying MBP immunofluorescence; the paper is not being cited to establish MBP as a marker of myelination. (LLM, conf=0.92)
plainconditional- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“we used an ultra high field 11.7 T scanner with a 3D DW-GRASE sequence that minimized distortion while allowing for a 12-fold acceleration compared to spin echo imaging (Aggarwal et al., 2010).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Partly supportedcites Aggarwal et al., 2010 sourceExperimental — accuracy not yet measured
Materials and Methods (DTI acquisition and Pulse sequence details; page unavailable) describe an 11.7 T instrument and a 3D DW-GRASE sequence. The Results (Comparison of GRASE and spin echo DTI; page unavailable) report no discernible geometric distortion relative to multiple spin echo imaging, but a threefold—not 12-fold—reduction in scan time. (LLM, conf=0.93)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“While temperature and fixation affect diffusivity values, and ADC in fixed tissue is significantly lower than in vivo (Richardson et al., 2014), anisotropy values are comparable between in vivo and ex vivo samples (Sun et al., 2001, 2003; Zhang et al., 2012).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Partly supportedcites Richardson et al., 2014 sourceExperimental — accuracy not yet measured
The Introduction states that temperature differences greatly affect measured diffusion coefficients, and DTI (Experiment A) reports lower axial diffusivity in fixed than viable isolated tissue. But the study compares fixed tissue with viable isolated tissue, not directly with in vivo samples, and the cited passages do not establish a significantly lower overall ADC. The anisotropy clause is attributed to other sources and is not part of this verdict. (LLM, conf=0.84)
- Not matchedcites Sun et al., 2001, 2003
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- Partly supportedcites Zhang et al., 2012 sourceExperimental — accuracy not yet measured
The Discussions report no significant difference between in vivo and ex vivo fractional anisotropy in the normal mouse corpus callosum. But they also report significantly different values in the demyelinated corpus callosum, so the unqualified claim that anisotropy is comparable across samples overextends the finding. (LLM, conf=0.88)
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“First, while it appears that we have labeled the major myelinated WM tracts of the mouse brain, there may be areas that are not fully captured due to incomplete antibody penetration or uneven labeling, which have been reported by other groups using intact brain approaches (Epp et al., 2015; Li et al., 2015).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Could not verifycites Epp et al., 2015 sourceExperimental — accuracy not yet measured
The citing authors’ statement about which tracts they labeled is their own; the point attributed to Epp et al. is the reported risk of incomplete antibody penetration or uneven labeling in intact-brain approaches. The Materials and Methods section describes antibody incubation, but neither it nor the passages found by searching for penetration and uneven labeling report that problem (page unavailable). (LLM, conf=0.72)
- Not matchedcites Li et al., 2015
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Could not verify
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In contrast, regions where clearly diverging fascicles could be observed in the CLARITY sample had lower linear anisotropy (Cl=0.35 ± 0.024) and also lower FA-MBP correlations (ρ=0.185, P=0.388, n=24 regions).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
plaincorrelationaln = 24- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Rests on an assumption: MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
- Rests on an assumption: The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
- Rests on an assumption: The ex vivo DTI scanning and CLARITY hydrogel perfusion do not alter the relationship between MBP and DTI metrics.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Specifically, mean MBP in the genu and splenium were both higher than in the body”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Rests on an assumption: MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
- Rests on an assumption: The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“we found that there were significant positive correlations for the genu (P < 0.0005) and splenium (P < 0.0005), but not for the body (P=0.138; Fig. 5D).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Rests on an assumption: MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
- Rests on an assumption: The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
In the paper
“The stria medullaris is a WM tract within the thalamus that carries axons from the septum to the habenula (Watson et al., 2012).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.5. Within-tract analysis of stria medullaris and corpus callosum
What the sources say
- Not checked — no full text of the sourcecites Watson et al., 2012 source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“This was unexpected for the scalar metric RD, which is considered to be sensitive to the degree of myelination and thus was expected to provide a significant negative correlation (Song et al., 2002; Sun et al., 2006a, 2006b).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 3. Results > 3.3. CLARITY MBP correlates with DTI-derived FA measures
What the sources say
- Not checked — no full text of the sourcecites Song et al., 2002 source
- Not matchedcites Sun et al., 2006a, 2006b
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
A claim is checked only against the cited paper’s full text, never its abstract, and none is available for its sources.
In the paper
“Diffusion tensor imaging (DTI) is the most commonly employed diffusion model and utilizes the magnitude and orientation of anisotropic water diffusion to estimate the WM structure of brain tissue, thereby producing maps of macroscopic axonal organization in the brain (Basser and Pierpaoli, 1996; Le Bihan et al., 2001; Mori and Zhang, 2006).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A claim is checked only against the cited paper’s full text, never its abstract, and none is available for its sources.
In the paper
“Diffusion-weighted MRI is the only noninvasive method available for mapping fiber architecture of tissue in vivo (Le Bihan et al., 1986; Basser et al., 1994)”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 1. Introduction
What the sources say
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Common sources of noise for in vivo neuroimaging include head motion, cardiac pulses, and arousal, which we have excluded by imaging ex vivo.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
A claim is checked only against the cited paper’s full text, never its abstract, and none is available for its sources.
In the paper
“we chose to label MBP as a putative biological index for myelination because it is the predominant protein within CNS myelin, representing as much as 30% of the protein in myelin (Baumann and Pham-Dinh, 2001) and is essential to myelin formation (Boggs, 2006).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A claim is checked only against the cited paper’s full text, never its abstract, and none is available for its sources.
In the paper
“Indeed, diffusion anisotropy is present even when compact myelin is completely absent, although it is significantly reduced (Roach et al., 1985; Watanabe et al., 2010).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Compared to the other WM ROIs, this subset tended to have larger volumes, which may minimize a potential partial volume effect that may have confounded the RD-MBP association in smaller WM tracts.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
In the paper
“Additionally, some of the RD-myelination association evidence is based on studies involving myelin damage to the mouse optic nerve (Song et al., 2003, 2006), which may differ from WM tracts within brain tissue.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Not matchedcites Song et al., 2003, 2006
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Not matched
In the paper
“The association of RD to myelination has been previously studied in some of these same WM structures, such as the corpus collosum and anterior commissure (Song et al., 2002), however, many other WM structures analyzed here have not been examined previously.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Not checked — no full text of the sourcecites Song et al., 2002 source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Another point of consideration is that the liquid hydrogel may have changed diffusivity during DTI in some unpredictable way. While we have not seen evidence for uneven labeling or altered diffusivity in our samples, we cannot rule these out as possibilities.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The relationship between FA and myelin is complex and cannot be resolved using this correlational approach alone.”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
In the paper
“Also, the percentage of myelinated axons and myelin sheath thickness are not necessarily indexed by MBP expression (Shine et al., 1992).”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
What the sources say
- Not checked — no full text of the sourcecites Shine et al., 1992 source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
The paper relies on this without support.
MBP immunofluorescence intensity is linearly proportional to myelin density across WM regions.
“MBP fluorescence intensity values ranged from 0–4095 and mean intensity values within ROIs were used for comparisons.”
The paper relies on this without support.
The CLARITY clearing and immunolabeling procedures do not differentially affect MBP detection across different WM regions.
“We performed the CLARITY procedure as previously described (Chung et al., 2013; Tomer et al., 2014; Yang et al., 2014) with custom modifications in order to maximize antibody penetration and minimize tissue expansion .”
The paper relies on this without support.
The ex vivo DTI scanning and CLARITY hydrogel perfusion do not alter the relationship between MBP and DTI metrics.
“In order to test the feasibility of CLARITY following DTI, we first tested three thy1-eYFP-H mice that underwent ∼18 h of ex vivo DTI in a high-field scanner.”
The paper relies on this without support.
Additional factors such as fiber architecture, axonal diameter, glial cells, and inflammation also contribute to diffusion anisotropy.
“However, additional factors such as fiber architecture, axonal diameter, glial cells, and inflammation also contribute to diffusion anisotropy.”
The paper relies on this without support.
The spatial resolution of diffusion MRI is low, typically on the order of 1–2 mm in the human brain in vivo, while myelinated axons are characterized on a micron scale.
“The spatial resolution of diffusion MRI is low, typically on the order of 1–2 mm in the human brain in vivo, while myelinated axons are characterized on a micron scale.”
The paper relies on this without support.
Diffusion anisotropy carries microscopic anatomical information, but this information is averaged over the large volume of an imaging voxel.
“While diffusion anisotropy does carry microscopic anatomical information, this information is averaged over the large volume of an imaging voxel.”
The paper relies on this without support.
None of the prior studies utilized a 3D-IHC approach for making comparisons to diffusion measures.
“None of the prior studies utilized a 3D-IHC approach for making comparisons to diffusion measures.”
The paper relies on this without support.
DTI remains a powerful method for noninvasively estimating WM structure in the brain with high sensitivity, but low specificity.
“Nonetheless, DTI remains a powerful method for noninvasively estimating WM structure in the brain with high sensitivity, but low specificity.”
The paper relies on this without support.
WM in the brain has a very complex structure that cannot be accurately estimated using a single diffusion tensor model, such as DTI.
“WM in the brain has a very complex structure that cannot be accurately estimated using a single diffusion tensor model, such as DTI.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“Studies have reported that FA from the DTI model was higher in more organized axonal bundles and showed its association with myelin basic protein (MBP).51”
From A spatial imaging-transcriptomics paradigm for deciphering the molecular basis of microscopic MRI in the normal brain and Alzheimer’s disease (2025) · cites this paper as [51]
Statistics recomputed
Every reported test result, recomputed from its test statistic and degrees of freedom. A result shows as not adding up only once an independent check has confirmed it.
- Consistentχ²(1) = 7.74reported p < .01 · recomputed p = .005
“Χ2(1)=7.74, p < .01”
- Consistentχ²(1) = 0.27reported p = .61 · recomputed p = .603
“Χ2(1)=0.27, p = .61”
- Consistentχ²(1) = 0.05reported p = .83 · recomputed p = .823
“Χ2(1)=0.05, p = .83”
- Consistentχ²(1) = 12.56reported p < .0005 · recomputed p < .001
“Χ2(1)=12.56, p < .0005”
- Consistentχ²(1) = 10.54reported p < .005 · recomputed p = .001
“Χ2(1)=10.54, p < .005”
- Consistentχ²(1) = 10.62reported p < .005 · recomputed p = .001
“Χ2(1)=10.62, p < .005”
- Consistentχ²(1) = 0.804reported p = .37 · recomputed p = .370
“Χ2(1)=0.804, p = .37”
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > 4. Discussion
“3DISCO (Ertürk et al., 2012)”
§ The role of myelination in measures of white matter integrity: Combination of diffusion tensor imaging and two-photon microscopy of CLARITY intact brains > References
“Ertürk A Becker K Jährling N Mauch CP Hojer CD Egen JG 1983 Three-dimensional imaging of solvent-cleared organs using 3DISCO”
Citations
Cites
Jenkinson, M., Bannister, P., Brady, M., & Smith, S. (2002) · NeuroImage · doi:10.1006/nimg.2002.1132 · Cited by 11,888
3D Slicer as an image computing platform for the Quantitative Imaging Network
Fedorov, A., Beichel, R., Kalpathy–Cramer, J., Finet, J., Fillion‐Robin, J., Pujol, S., Bauer, C., Jennings, D. L., Fennessy, F., Milan Sonka, A., et al. (2012) · Magnetic Resonance Imaging · doi:10.1016/j.mri.2012.05.001 · Cited by 9,095
MR diffusion tensor spectroscopy and imaging
Basser, P. J., Mattiello, J., & LeBihan, D. (1994) · Biophysical Journal · doi:10.1016/s0006-3495(94)80775-1 · Cited by 5,970
Genome-wide atlas of gene expression in the adult mouse brain
Lein, E. S., Hawrylycz, M. J., Ao, N., Ayres, M., Bensinger, A., Bernard, A., Boe, A. F., Boguski, M. S., Brockway, K. S., Byrnes, E. J., et al. (2007) · Nature · doi:10.1038/nature05453 · Cited by 5,572
The basis of anisotropic water diffusion in the nervous system—A technical review.
Beaulieu, C. (2002) · NMR in Biomedicine · doi:10.1002/nbm.782 · Cited by 4,628
Basser, P. J., & Pierpaoli, C. (1996) · Journal of Magnetic Resonance, Series B · doi:10.1006/jmrb.1996.0086 · Cited by 4,306
Le Bihan, D., Breton, E., Lallemand, D., Grenier, P., Cabanis, E., & Laval-Jeantet, M. (1986) · Radiology · doi:10.1148/radiology.161.2.3763909 · Cited by 3,912
Diffusion tensor imaging: Concepts and applications
Le Bihan, D., Mangin, J., Poupon, C., Clark, C. A., Pappata, S., Molko, N., & Chabriat, H. (2001) · Journal of Magnetic Resonance Imaging · doi:10.1002/jmri.1076 · Cited by 3,547
A mesoscale connectome of the mouse brain
Oh, S. W., Harris, J. A., Ng, L., Winslow, B., Cain, N., Mihalas, S., Wang, Q., Lau, C., Kuan, L., Henry, A. M., et al. (2014) · Nature · doi:10.1038/nature13186 · Cited by 2,612
Dysmyelination revealed through MRI as increased radial (but unchanged axial) diffusion of water
Song, S.-K., Sun, S.-W., Ramsbottom, M. J., Chang, C., Russell, J., & Cross, A. H. (2002) · NeuroImage · doi:10.1006/nimg.2002.1267 · Cited by 2,582
Cited by
Sorted by citations
White Matter Plasticity in the Adult Brain
Sampaio‐Baptista, C., & Johansen‐Berg, H. (2017) · Neuron · doi:10.1016/j.neuron.2017.11.026 · Cited by 432
“…and whole-brain immunolabeling along with the diffusion tensor model (DTI) demonstrated that myelin basic protein (MBP) correlates with FA in WM regions with coherent fiber orientations and low fiber dispersion, but not in areas of crossing or complex fiber architecture (Chang et al., 2017).”
BackgroundCommon genetic variation influencing human white matter microstructure
Zhao, B., Li, T., Yang, Y., Wang, X., Luo, T., Shan, Y., Zhu, Z., Xiong, D., Hauberg, M. E., Bendl, J. M., et al. (2021) · Science · doi:10.1126/science.abf3736 · Cited by 222
“Aspects of this connectivity can be estimated through measurements of cellular microstructure (7).”
BackgroundDesign and Validation of Diffusion MRI Models of White Matter
Jelescu, I. O., & Budde, M. D. (2017) · Frontiers in Physics · doi:10.3389/fphy.2017.00061 · Cited by 240
“Many of the whole-specimen imaging techniques developed for local neuron connectomic studies [139–141] have demonstrated utility when coupled with diffusion MRI [142].”
Leclercq, S., Le Roy, T., Furgiuele, S., Coste, V., Bindels, L. B., Leyrolle, Q., Neyrinck, A. M., Quoilin, C., Amadieu, C., Petit, G., et al. (2020) · Cell Reports · doi:10.1016/j.celrep.2020.108238 · Cited by 178
“In the mouse brain, FA correlates positively with the myelin basic protein, a marker for myelination (Chang et al., 2017), and it has been shown in AUD patients that reduced FA in the corpus callosum was compatible with sustained demyelination and glial immune reaction (De Santis et al.”
BackgroundLazari, A., & Lipp, I. (2021) · NeuroImage · doi:10.1016/j.neuroimage.2021.117744 · Cited by 176
Metadata
- Authors
- Chang, E. H., Argyelan, M., Aggarwal, M., Chandon, T.-S. S., Karlsgodt, K. H., Mori, S., & Malhotra, A. K.
- Affiliations
- Northwell Health (US)
- Johns Hopkins University (US)
- Kennedy Krieger Institute (US)
- Corresponding author
- Chang, Eric H.
- Venue
- NeuroImage
- Publisher
- Elsevier BV
- Published
- 2017-02
- Volume
- 147
- Pages
- 253-261
- ISSN
- 1053-8119 (ISSN-L)
- PMC
- PMC5560594
- OpenAlex
- W2563000655
- Semantic Scholar
- aa30e8c1e8b1d8d5eeafb64fca929a49ee935650
- Cited by
- 186
- References
- 66
- Citations by year
- 2026: 16 · 2025: 27 · 2024: 13 · 2023: 14 · 2022: 17 · 2021: 27 · 2020: 19 · 2019: 17 · 2018: 21 · 2017: 13
- FWCI
- 13.66
- Type
- journal article
- Language
- en
- Open access
- green
- escholarship.org
- Locations
- NeuroImage (publishedVersion)
- PubMed (publishedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- PubMed Central (submittedVersion)
- Funders
- Center for Intervention Development and Applied Research (P50MH080173)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Advanced Neuroimaging Techniques and Applications
- Life Sciences › Neuroscience › Cognitive Neuroscience › Functional Brain Connectivity Studies
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Advanced MRI Techniques and Applications
- Keywords
- myelination, white matter integrity, diffusion tensor imaging, myelin basic protein, fractional anisotropy, radial diffusivity, fiber dispersion, mouse brain, ex vivo brain imaging
- MeSH terms
- Animals, Fluorescent Antibody Technique, Male, Mice, Inbred C57BL, Myelin Sheath, Anisotropy, Microscopy, Fluorescence, Multiphoton, Mice, Diffusion Tensor Imaging, White Matter
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
A language model drew the claims from the paper on 2026-10-24, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-24 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Cited sources are checked with a language model’s help (gpt-6-sol).
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
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