A novel method for evaluation of capillarity in human skeletal muscles from confocal 3D images

Janáček, Jiřı́; Cvetko, Erika; Kubínová, Lucie; Travnik, Ludvik; Eržen, Ida

doi:10.1016/j.mvr.2010.11.012

Cited by 23 publications

(21 citation statements)

References 40 publications

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“…The fakir probes were randomly oriented in 3-dimentions, thereby accounting for tissue orientation bias. (Kubinova, 1998; Kubinova and Janacek, 1998, 2001; Kubinova et al, 2001; Kubinova et al, 2003; Janacek et al, 2011)…”

Section: Methodsmentioning

confidence: 99%

Idiopathic Microscopic Colitis of Rhesus Macaques: Quantitative Assessment of Colonic Mucosa

Ardeshir

Oslund

Ventimiglia

et al. 2013

The Anatomical Record

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Idiopathic chronic diarrhea (ICD) is a common cause of morbidity and mortality among juvenile rhesus macaques. While lesions may be absent at colonoscopy, the histopathologic evaluation of the biopsy specimens is consistent with human macroscopic colitis (MC). In this study, we developed an isotropic uniform random sampling method to evaluate macroscopic and microscopic changes and applied it on proximal ascending colon in monkeys. Colonic tissue and peripheral blood specimens were collected from six MC and six control juvenile macaques at necropsy. Uniform random samples were collected from the colon using punch biopsy tools. The volume of epithelium and lamina propria were estimated in thick (25 µm) sections using point probes and normalized to the area of muscularis mucosae. Our data suggests a significant increase of the Vs of the lamina propria (1.9 fold, p=0.02) and epithelium (1.4 fold, p=0.05) in subjects with MC. The average colonic surface mucosa area in the MC monkeys increased 1.4 fold over the controls (p=0.02). The volume of the proximal colon in animals with MC showed a 2.4 fold increase over the non-diarrhea control monkeys (p=0.0001). Cytokine, chemokine, and growth factor levels in peripheral blood were found to be correlated with the volume estimate of the lamina propria and epithelium. We found that ICD in macaques has features which simulates human MC and can be used as a spontaneous animal model for human MC. Furthermore, this developed sampling method can be used for unbiased evaluation of therapeutics in clinical trials of this animal model.

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Section: Methodsmentioning

confidence: 99%

Idiopathic Microscopic Colitis of Rhesus Macaques: Quantitative Assessment of Colonic Mucosa

Ardeshir

Oslund

Ventimiglia

et al. 2013

The Anatomical Record

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“…Local parameters were determined by manual analysis of 5-μm-thin tissue sections, where capillaries were double stained with fluorescein and rhodaminelabelled lectin, and muscle fibre outlines were unstained. Although staining of the basal lamina or the sarcolemma of muscle fibres with anti-fibronectin, anti-laminin or anti-desmin antibodies can be very useful for image analysis [Hantaï et al, 1984;Kubínová et al, 2001;Čebašek et al, 2004;Janáček et al, 2009;Meunier et al, 2010;Janáček et al, 2011;Talmadge et al, 2014], applying only the phasecontrast microscopy for visualization of muscle fibre outlines improves visibility of capillaries and facilitates manual tracing of capillaries.…”

Section: Muscle Fibre Morphometry and Capillary Characteristicsmentioning

confidence: 99%

Changes in the Capillarity of the Rat Extensor Digitorum Longus Muscle 4 Weeks after Nerve Injury Studied by 2D Measurement Methods

Čebašek¹,

Ribarič

2016

Cells Tissues Organs

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We have previously shown by 3D study that 2 weeks after nerve injury there was no change in the length of capillaries per muscle fibre length in rat extensor digitorum longus muscle (EDL). The primary goal of the present 2D study was to determine the capillarity of rat EDL 4 weeks after various modes of nerve injury. Additionally, we wished to calculate the same capillary/fibre parameters that were used in our 3D stereological study. EDL muscles derived from denervated (4 weeks after nerve injury), re-innervated (4 weeks after two successive nerve crushes) and age-matched controls from the beginning (CON-1) and the end (CON-2) of the experiment were analysed in two ways. Global indices of capillarity, such as capillary density (CD) and capillary/fibre (C/F) ratio, were determined by automatic analysis, local indices as the number (CAF) and the length of capillaries around individual muscle fibres (Lcap) in relation to muscle fibre size were estimated manually by tracing the muscle fibre outlines and the transversally and longitudinally cut segments of capillaries seen in 5-µm-thin muscle cross sections. Four weeks after both types of nerve injury, CD increased in comparison to the CON-2 group (p < 0.001) due to atrophied muscle fibres in denervated muscles and probably proliferation of capillaries in re-innervated ones. Higher C/F, CAF (both p < 0.001) and Lcap (p < 0.01) in re-innervated than denervated EDL confirmed this assumption. Calculated capillary/fibre parameters were comparable to our previous 3D study, which strengthens the practical value to the adapted 2D method used in this study.

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“…Moreover, to our knowledge it is practically impossible to control the extent of autolysis. In Janáček et al (2011) we describe in detail the methodological problems that needed to be solved before we could get a good staining of capillaries through 100 µm muscle section. Further, although UAE lectin is an excellent marker for capillaries in human skeletal muscles, it did not function in several human autopsy muscle samples.…”

Section: Discussionmentioning

confidence: 99%

“…To prevent tissue deformation the distance between the slide and the cover slip was maintained by two layers made by the ImgEdge pen (Dako, Glostrup, Denmark) (see Janáček et al, 2011).…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…To avoid this estimation error we applied the method for estimating characteristics of the capillary network in 3D, introduced in our previous publications (Kubínová et al, 2001;Eržen et al, 2011;Janáček et al, 2011). The method requires thick tissue sections from which a complete series of optical sections is captured by a confocal microscope.…”

Section: Introductionmentioning

confidence: 99%

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The Capillary Pattern in Human Masseter Muscle During Ageing

et al. 2013

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The effect of ageing on the capillary network in skeletal muscles has produced conflicting results in both, human and animals studies. Some of the inconsistencies are due to non-comparable and biased methods that were applied on thin transversal sections, especially in muscles with complicated morphological structures, such as in human masseter muscle. We present a new immunohistochemical method for staining capillaries and muscle fibres in 100 µm thick sections as well as novel approach to 3D visualization of capillaries and muscle fibres. Applying confocal microscopy and virtual 3D stereological grids, or tracing capillaries in virtual reality, length of capillaries within a muscle volume or length of capillaries adjacent to muscle fibre per fibre length, fibre surface or fibre volume were evaluated in masseter muscle of young and old subjects by an unbiased approach. Our findings show that anatomic capillarity is well maintained in masseter muscle in old subjects; however, vascular remodelling occurs with age, which could be a response to changed muscle function and age-related muscle fibre type transformations.

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A novel method for evaluation of capillarity in human skeletal muscles from confocal 3D images

Cited by 23 publications

References 40 publications

Idiopathic Microscopic Colitis of Rhesus Macaques: Quantitative Assessment of Colonic Mucosa

Idiopathic Microscopic Colitis of Rhesus Macaques: Quantitative Assessment of Colonic Mucosa

Changes in the Capillarity of the Rat Extensor Digitorum Longus Muscle 4 Weeks after Nerve Injury Studied by 2D Measurement Methods

The Capillary Pattern in Human Masseter Muscle During Ageing

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