Cryogenic contrast-enhanced microCT enables nondestructive 3D quantitative histopathology of soft biological tissues

Maes, Arne; Pestiaux, Camille; Marino, Alice; Balcaen, Tim; Leyssens, Lisa; Vangrunderbeeck, Sarah; Pyka, Grzegorz; Borggraeve, Wim M. De; Bertrand, Luc; Beauloye, Christophe; Horman, Sandrine; Wevers, Martine; Kerckhofs, Greet

doi:10.1038/s41467-022-34048-4

Cited by 22 publications

(29 citation statements)

References 82 publications

(100 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sample size is consistent with recent studies using XRM/MicroCT for visualizing biological structures and are generally n = 3 per group in this study 74 . Statistical methods were not used to re-calculate or predetermine sample sizes in any experiments.…”

Section: Methodssupporting

confidence: 86%

Gold nanoparticle-enhanced X-ray microtomography of the rodent reveals region-specific cerebrospinal fluid circulation in the brain

et al. 2023

View full text Add to dashboard Cite

Cerebrospinal fluid (CSF) is essential for the development and function of the central nervous system (CNS). However, the brain and its interstitium have largely been thought of as a single entity through which CSF circulates, and it is not known whether specific cell populations within the CNS preferentially interact with the CSF. Here, we develop a technique for CSF tracking, gold nanoparticle-enhanced X-ray microtomography, to achieve micrometer-scale resolution visualization of CSF circulation patterns during development. Using this method and subsequent histological analysis in rodents, we identify previously uncharacterized CSF pathways from the subarachnoid space (particularly the basal cisterns) that mediate CSF-parenchymal interactions involving 24 functional-anatomic cell groupings in the brain and spinal cord. CSF distribution to these areas is largely restricted to early development and is altered in posthemorrhagic hydrocephalus. Our study also presents particle size-dependent CSF circulation patterns through the CNS including interaction between neurons and small CSF tracers, but not large CSF tracers. These findings have implications for understanding the biological basis of normal brain development and the pathogenesis of a broad range of disease states, including hydrocephalus.

show abstract

Section: Methodssupporting

confidence: 86%

Gold nanoparticle-enhanced X-ray microtomography of the rodent reveals region-specific cerebrospinal fluid circulation in the brain

et al. 2023

View full text Add to dashboard Cite

show abstract

“…The 16-bit slices (.tiff) were converted 8-bit slices (.jpg) using an in-house developed MatLab tool, which simultaneously windowed the histogram range to the dynamic range of the dataset and normalized the data to the last Hf-WD POM dataset 39 . In the normalization step, the grey values were matched between datasets for the two reference materials: the cortical bone of the cochlear capsule and the parafilm foil.…”

Section: Methodsmentioning

confidence: 99%

“…The objective of this study was to obtain simultaneous 3D visualization of the mineralized and soft-tissue intracochlear structures, which are relevant for CI surgery, in fresh human cadaveric cochleae in a fully nondestructive manner. To achieve this, we made use of a promising CESA, which has been recently introduced for soft tissue visualization on CECT: a 1:2 hafnium-substituted Wells–Dawson polyoxometalate (K 16 [Hf(α 2 -P 2 W 17 O 61 ) 2 ]·19H 2 O), further referred to as Hf-WD POM 36 – 39 . Similar to phosphotungstic acid, Hf-WD POM has a binding affinity for fibrin, collagen and fibers of connective tissue.…”

Section: Introductionmentioning

confidence: 99%

Human cochlear microstructures at risk of electrode insertion trauma, elucidated in 3D with contrast-enhanced microCT

Starovoyt

Pyka

Putzeys

et al. 2023

Sci Rep

Self Cite

View full text Add to dashboard Cite

Cochlear implant restores hearing loss through electrical stimulation of the hearing nerve from within the cochlea. Unfortunately, surgical implantation of this neuroprosthesis often traumatizes delicate intracochlear structures, resulting in loss of residual hearing and compromising hearing in noisy environments and appreciation of music. To avoid cochlear trauma, insertion techniques and devices have to be adjusted to the cochlear microanatomy. However, existing techniques were unable to achieve a representative visualization of the human cochlea: classical histology damages the tissues and lacks 3D perspective; standard microCT fails to resolve the cochlear soft tissues; and previously used X-ray contrast-enhancing staining agents are destructive. In this study, we overcame these limitations by performing contrast-enhanced microCT imaging (CECT) with a novel polyoxometalate staining agent Hf-WD POM. With Hf-WD POM-based CECT, we achieved nondestructive, high-resolution, simultaneous, 3D visualization of the mineralized and soft microstructures in fresh-frozen human cochleae. This enabled quantitative analysis of the true intracochlear dimensions and led to anatomical discoveries, concerning surgically-relevant microstructures: the round window membrane, the Rosenthal’s canal and the secondary spiral lamina. Furthermore, we demonstrated that Hf-WD POM-based CECT enables quantitative assessment of these structures as well as their trauma.

show abstract

“…All the other datasets were normalized to that one using a second in-house developed MATLAB script with the borosilicate bead and the sample holder as reference materials. MATLAB scripts are available on github ( 27 , 28 ).…”

Section: Methodsmentioning

confidence: 99%

3D histopathology of stenotic aortic valve cusps using ex vivo microfocus computed tomography

Pestiaux¹,

Pyka²,

Quirynen³

et al. 2023

Front. Cardiovasc. Med.

Self Cite

View full text Add to dashboard Cite

BackgroundCalcific aortic stenosis (AS) is the most prevalent heart valve disease in developed countries. The aortic valve cusps progressively thicken and the valve does not open fully due to the presence of calcifications. In vivo imaging, usually used for diagnosis, does not allow the visualization of the microstructural changes associated with AS.MethodsEx vivo high-resolution microfocus computed tomography (microCT) was used to quantitatively describe the microstructure of calcified aortic valve cusps in full 3D. As case study in our work, this quantitative analysis was applied to normal-flow low-gradient severe AS (NF-LG-SAS), for which the medical prognostic is still highly debated in the current literature, and high-gradient severe AS (HG-SAS).ResultsThe volume proportion of calcification, the size and number of calcified particles and their density composition was quantified. A new size-based classification considering small-sized particles that are not detected with in vivo imaging was defined for macro-, meso- and microscale calcifications. Volume and thickness of aortic valve cusps, including the complete thickness distribution, were also determined. Moreover, changes in the cusp soft tissues were also visualized with microCT and confirmed by scanning electron microscopy images of the same sample. NF-LG-SAS cusps contained lower relative amount of calcifications than HG-SAS. Moreover, the number and size of calcified objects and the volume and thickness of the cusps were also lower in NF-LG-SAS cusps than in HG-SAS.ConclusionsThe application of high-resolution ex vivo microCT to stenotic aortic valve cusps provided a quantitative description of the general structure of the cusps and of the calcifications present in the cusp soft tissues. This detailed description could help in the future to better understand the mechanisms of AS.

show abstract

Cryogenic contrast-enhanced microCT enables nondestructive 3D quantitative histopathology of soft biological tissues

Cited by 22 publications

References 82 publications

Gold nanoparticle-enhanced X-ray microtomography of the rodent reveals region-specific cerebrospinal fluid circulation in the brain

Gold nanoparticle-enhanced X-ray microtomography of the rodent reveals region-specific cerebrospinal fluid circulation in the brain

Human cochlear microstructures at risk of electrode insertion trauma, elucidated in 3D with contrast-enhanced microCT

3D histopathology of stenotic aortic valve cusps using ex vivo microfocus computed tomography

Contact Info

Product

Resources

About