DNA ploidy analysis in histopathology. Morphometry and DNA cytometry reproducibility conditions and clinical applications.
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Cytological Techniques↗
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Results for “morphometry”
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The use of simple image analysers in lung morphometry.
We have used several commercially available image analysers to measure the inter-alveolar wall distance on histologic sections of animal and human lungs. Inter- and intra-observer variations using these machines are small, and very similar results are obtained using different machines. The recorded result depends on the magnification used since higher resolution produces more intercepts. The plane in which the measurement is made affects the result, representing the difference in shrinkage from fixed tissue to stained slides in the horizontal and vertical planes. Machine-measured inter-alveolar wall distances are smaller than those measured by humans because of the spurious intercepts made by the image analysers. However, the measurements are highly correlated and human-measured inter-alveolar wall distances can be closely predicted from the machine-measured ones. The number of alveoli per unit volume is less well predicted from the average inter-alveolar wall distance. Image analysers may possibly play a useful role in making similar simple measurements in other organs.
Animals↗
Comparison of two subsampling methods for electron microscopic morphometry.
The purpose of the present paper is twofold. Firstly, to compare the accuracy per unit cost achieved by two different methods of subsampling micrographs from sections of a material for stereology when the sections cannot be analysed as a whole at the required magnification. Secondly, to illustrate, by means of real data, the application of some of the methods and formulae proposed in the companion paper (Cruz-Orive & Weibel, 1981) for estimating ratios at the electron microscopic level. The final estimates of a same ratio obtained by either subsampling method (namely systematic (SQ) and systematic area-weighted quadrats (SAWQ)) agreed in the mean and they were about equally precise. The former fact indicates that the new SAWQ method is at least as reliable as the SQ method as far as bias is concerned. The latter result is a consequence of the well-known fact that subsampling is relatively unimportant in two-stage sampling. Yet, SAWQ subsampling enjoys definite practical advantages over other subsampling methods in certain situations.
Anatomy↗
The use of touch-sensitive screen in interactive morphometry.
We propose the use of a touch-sensitive screen as a tool to facilitate acquisition of data both in point counting and morphometric planimetry. It obviates the use of a 'correspondence image' required with many planimetric devices in which the means for data acquisition and display are separate. The touch-sensitive screen offers operational advantages over the light pen. The characteristics of this new graphical input device, its assembly and basic operational considerations are introduced and discussed.
Computers↗
Surface exchange area of the porcine placenta: morphometry of anisotropic interdigitating microvilli.
The pig has a diffuse epitheliochorial placenta. The foeto-maternal surface area (Sv(F), Sv(M)) is enlarged at three levels: level I: macroscopic folds of uterine mucosa covered by foetal membranes; level II: light microscopic folds of uterine epithelium covered by trophoblastic epithelium; level III: ultrastructural formation of both maternal and foetal microvilli in an interdigitating pattern. This study presents a morphometric method, using intersection counting, to estimate the surface area of these interdigitating, highly oriented microvilli at level III, i.e. the 'true' surface exchange area. The method is applied to different stages in pregnancy. The epithelial surfaces are enlarged by a factor 8-11 due to the formation of microvilli. This factor varies during pregnancy, being lower early in gestation and before delivery, and for the first time it is shown to be significantly different for mother and foetus. At level III the foetal surface of placenta is larger than the maternal indicating that the two opposing surfaces are not congruent at the ultrastructural level and that membrane-linked placental transport may be different in mother and foetus. A three-compartment model is suggested for placental exchange with maternal, intermicrovillous and foetal compartments.
Animals↗
Midfacial and mandibular morphometry of children with Class II and Class III malocclusions.
There is insufficient evidence in conventional cephalometric analysis of the actual sites of putative maxillofacial change in Class II and Class III malocclusions. The purpose of this study was to provide more information about the morphological characteristics of the midfacial complex and mandible in children with Class II or III malocclusions. Seventy children with Class II, division 1 malocclusion and 70 children with Class III malocclusion were compared with 70 children with normal occlusion. This study was conducted to carry out geometric morphometric assessments to localize alterations using Procrustes analysis and thin-plate spline analysis. Procrustes analysis indicated the midfacial and mandibular morphologies differed between normal occlusion subjects and subjects with Class II or Class III malocclusion (P<0.0001). The deformations in subjects with Class II malocclusion may represent a developmental elongation of the palatomaxillary complex and a shortening of the mandible anteroposteriorly, which leads to the appearance of a protruding midface and retruding mandibular profile. In contrast, the deformations in subjects with Class III malocclusion may represent a developmental shortening of the palatomaxillary complex and elongation of the mandible anteroposteriorly, which leads to the appearance of a retrognathic midface and prognathic mandibular profile.
Cephalometry↗
The morphology and morphometry of the normal human tibialis anterior muscle.
The light microscopic appearance of the human tibialis anterior muscle is described based on conchotome biopsy specimens from seven healthy volunteers and 20 patients who presented with myalgia but who had no evidence of neuromuscular disease. The morphometric characteristics of these normal muscles are documented and the similarities and differences between the appearances of the tibialis anterior and other muscles discussed.
Adult↗
Muscle morphometry in motor neuron disease.
It has previously been suggested that the pathological abnormalities seen in muscle biopsies from patients with motor neuron disease (MND) are of predictive value in relation to the rate of progression of the disease. In this study, quadriceps muscle biopsies from 19 patients with MND and 20 age matched controls were prepared for histochemistry and analysed morphometrically. Pathological features of denervation and reinnervation were observed in all MND patients although considerable variation between patients was noted. Motor neuron disease biopsies also showed increased connective tissue, an increased variation in fibre size, and a random fibre type distribution. Several of these abnormalities were more severe in female patients. Many of these 'abnormalities' were also frequent, albeit to a milder degree, in control biopsies and emphasize the need for age matched controls. The morphometric data was not related to the age of the patient, disease duration, type of MND or muscle strength, thus suggesting that the progression and severity of MND and its prognosis cannot be judged on the basis of quadriceps muscle pathology alone.
Adult↗
Early unsuspected neuron and axon terminal loss in scrapie-infected mice revealed by morphometry and immunocytochemistry.
Neuronal loss is often quoted as an element of the pathology of the transmissible spongiform encephalopathies, but few data are published. To determine whether neuronal loss is a salient feature of murine scrapie, and whether there is a relationship with the other hallmark lesions of scrapie we compared the numbers of neurons, severity of vacuolation, axonal bouton density and distribution of prion protein (PrP) in the dorsal lateral geniculate nucleus (dLGN) following intraocular infection of C57BL/FaBtDk mice with ME7 scrapie. This route of infection limits the initial spread of infection to the retinal efferents, thus directing infectivity and subsequent pathological changes to the dLGN which is a major projection of the optic nerve. Morphometric assessment of neuron number in the dLGN was made on semi-serial sections from five infected and five normal brain injected controls at four 50-day intervals during the incubation period, and on terminally affected mice. The number of neurons decreased from around 20,000 at 50 days to under 1000 in the terminal group. Significant loss was identified in individual mice at 150 days post-infection, coincident with the onset of vacuolation: neuron number was found to have an inverse relationship to the severity of vacuolation. Axonal boutons in the dLGN (demonstrated by synaptophysin immunolabelling) were reduced at 200 days, and virtually absent in terminal mice. The intensity of PrP immunostaining progressively increased from 150 days, and in a separate experiment PrP was detected from 175 days by polyacrylamide gel electrophoresis of brain extracts.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Hypoxia-inducible factor 1alpha/vascular endothelial growth factor axis in astrocytomas. Associations with microvessel morphometry, proliferation and prognosis.
Hypoxia-inducible factor (HIF)-1alpha is a transcription factor that promotes ischaemia-driven angiogenesis. The aim of this study was to determine the relation of HIF-1alpha to vascular endothelial growth factor (VEGF; an important angiogenic molecule in brain tumours), p53 expression, angiogenesis, proliferative potential and clinical outcome in a large series of diffuse astrocytomas. Expression of HIF-1alpha, VEGF, Ki-67 (a proliferation-associated marker) and p53 was determined immunohistochemically in 83 adult patients with supratentorial diffuse astrocytomas. Microvessels, highlighted by means of anti-CD34 immunohistochemistry, were enumerated with computer-assisted image analysis. Although HIF-1alpha and VEGF were expressed in the majority of cases, their levels increased significantly with increasing grade and proliferative potential. HIF-1alpha positively correlated with microvessel counts and VEGF with total vascular area and the presence of rounder vessel sections. There was a positive correlation of VEGF with p53 expression in astrocytomas and anaplastic astrocytomas. In univariate analysis, both VEGF and HIF-1alpha were associated with shortened survival in the entire cohort, but lost significance when grades II/III and grade IV were analysed separately. Multivariate analysis revealed that the combination of HIF-1alpha with grade was a significant prognostic indicator. HIF-1alpha expression may be used to refine the prognostic information provided by grade in patients with diffuse astrocytomas. Its adverse prognostic effect is most likely mediated by hypoxia, the driving force for HIF-1alpha accumulation.
Adolescent↗
Cell proliferation rate and nuclear morphometry in Roberts syndrome.
Roberts syndrome (RS) is a rare autosomal recessive disorder characterized primarily by symmetric reduction anomalies of all limbs, growth retardation and craniofacial abnormalities. Most RS patients are reported to present a typical abnormality of their constitutive heterochromatin, accompanied by abnormal cytological growth characteristics. We present an extremely severe case of an RS fetus, karyotypically documented, with a clinical presentation including growth deficiency, tetraphocomelia, frontal meningocele, craniofacial abnormalities and penile enlargement with hypospadias. Nuclear morphometrical analysis in tissues of various organs revealed a reduced nuclear size in RS as compared to normal controls, and statistically significant differences in morphometric parameters related to the nuclear shape. Immunohistochemical study of the same organs showed a reduced expression of proliferating cell nuclear antigen in the presented case, thus indicating a decreased cell proliferation rate in RS. Our results reconfirm previously reported findings in cultured fibroblasts of RS cases, thereby reinforcing on a histologic level, the hypothesis that reduced cell proliferation may be involved in the growth retardation and the reduction abnormalities observed in RS.
Abnormalities, Multiple↗
When morphometry meets genetics: inferring the phylogeography of Carabus solieri using Fourier analyses of pronotum and male genitalia.
Population differentiation is a crucial step in the speciation process and is therefore a central subject in studies of microevolution. Assessing divergence and inferring its dynamics in space and time generally require a wide array of markers. Until now however, most studies of population structure are based on molecular markers and those concerning morphological traits are more scarce. In the present work, we studied morphological differentiation among populations of the ground beetle Carabus solieri, and tested its congruence with genetic population structure. The shape of pronotum and aedeagus was assessed using Dual Axis Fourier Shape Analysis. manova on Fourier coefficients revealed highly significant morphological variation between populations and a similar geographical pattern of differentiation for both structures. On the whole, morphological and genetic patterns were also found to be congruent. Our analysis confirms the phylogeographical scenario proposing that two entities of C. solieri differentiated during the last glaciation events before recolonizing the actual range of the species. It also indicates a large introgression between the two differentiated entities in the centre of the range.
Analysis of Variance↗
Muscle fibre morphometry in three dog muscles of different functional purpose in different breeds.
Following a study of fibre population fibre size was determined in three muscles of different functional purpose in four breeds of dog with different locomotory capacities. Samples were taken from specific sites in the tibialis cranialis (extensor), flexor digitalis medialis (flexor) and pectineus (adductor) muscles of 27 dogs of four different breeds--German shepherd, Spanish greyhound, Spanish mastiff and Iberian hound--. Samples were subjected to histochemical analysis using ATPase (pH 9.4) and fibre size was assessed using an IBAS-2 Kontron image analyser.
Animals↗
Canine spinal cord neuron and axon myelin sheath morphometry.
This inedited morphometric study has been developed from healthy canine spinal cord neuron cytoplasm and nucleus, and white matter axonal myelin sheath, from cervical, thoracic and lumbar regions. For the morphometric study, the parameters were area, perimeter, maximum and minimum diameters and roundness for neurons and myelin thickness for axon. For each parameter, 300 neurons were analysed. The results revealed that lumbar neurons had the highest mean values for the analysed parameters, indicating the presence of large neurons in this region, with large axons as a result of myelin thickness, which is proportional to axon calibre. We conclude that these morphometric results can contribute for the establishment of normal patterns, for canine spinal cord cervical, thoracic and lumbar segments.
Animals↗
Ultrastructural morphometry of mammary gland in transgenic and non-transgenic rabbits.
The mammary gland of transgenic animals has been used for the production of recombinant proteins of therapeutic and nutraceutical use. The objective of this study was to compare the ultrastructure of transgenic and non-transgenic rabbit mammary gland tissue. New Zealand White transgenic rabbits were obtained by breeding non-transgenic rabbits with transgenic founder rabbits containing a whey acidic protein-human factor VIII (WAP-hFVIII) transgene integrated into their genome. Samples of mammary gland tissue from lactating rabbit females were isolated by surgical procedures. These samples were examined by optical and electron microscopy and photographs were taken. Measurements of ultrastructural organelles were made from digital images of the mammary cells. No differences were found in the cellular structure of mammary tissue, but significant differences t((0.001)) in the relative volume of mitochondria and vacuoles between transgenic and non-transgenic mammary gland epithelium were observed.
Animals↗
Gonadal response to psychosocial stress in male tree shrews (Tupaia belangeri) morphometry of testis, epididymis and prostate.
Male tree shrews (46 animals in all) were kept under distinct social conditions. Two kinds of controls were investigated: isolated animals as well as males out of female/male couples. Comparing these two groups only little isolation effect was observed. With the other individuals, initially isolated, dyads were formed which coexisted for 7 or 50 days. In a third group of dyads, after 50 days of coexistence social circumstances were aggravated for 5 more days by providing only half of the cage. All dyads established clear cut dominance relationship. Animals behaving dominantly developed no signs of gonadal regression rather that of stimulation concerning especially with the epididymis. In contrast, animals behaving as subordinates revealed profound regressive changes although differentiated with respect to any parameter, both morphological and conditional. In testis, chronical subordination (50 days, mainly 50 + 5 days) leads to a loss of organ weight, narrowing of seminiferous tubules and to enlarged spermatogonia nuclei. The case is similar in the epididymis: loss of organ weight, lowered epithelium and an enormously increased frequency of immature germ cells within tail segments. Whereas prostate weight drops continuously, striking histological regression occurs already after 7 days (extreme diminution of epithelial height and of epithelial cell nuclei).
Animals↗
Comparisons of sperm quality, morphometry and function among human sperm populations recovered via SpermPrep II filtration, swim-up and Percoll density gradient methods.
The purpose of this study was to compare the morphology/morphometry and fertilizing capacity of human spermatozoa recovered via swim-up method, Percoll density gradient method, and SpermPrep II filtration method. Thirty-three ejaculates were equally divided into 2 aliquots. Aliquot 1 was processed via the direct swim-up method, whereas aliquot 2 was filtered via a SpermPrep II column. The Percoll density gradient method was compared with the SpermPrep II method in a similar protocol using 43 ejaculates. Sperm populations recovered via the SpermPrep II filtration method showed significantly higher hypoosmotic swelling test results, acrosin profiles, and percentage of hyperactivated spermatozoa than sperm fractions recovered by the swim-up method. Furthermore, significant differences were found in most of sperm morphometric parameters between the above sperm populations. However, sperm fractions recovered via the SpermPrep II method did not show significantly different values for these same tests and for most of sperm morphometric parameters compared to the Percoll density gradient method. These results suggest that the SpermPrep II filtration and Percoll density gradient method are equally efficient in isolating sperm subpopulations with better functional parameters than the swimup method.
Cell Separation↗
Porcine congenital tremor type AI: spinal cord morphometry.
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Animals↗