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PubMed1993

Morphometry of human epidermis in vivo by real-time confocal microscopy.

Real-time confocal microscopy has brought substantial improvements to the imaging of the human skin in vivo. On early images, the stratum corneum could be distinguished from the living epidermis and the circulatory network of the superficial dermis. We have adapted the Tandem Scanning Microscope to obtain images of the living skin, showing thinner structures such as the stratum lucidum and the dermo-epidermal junction, both of which are essential markers for micron-order measurements of the thickness of the stratum corneum and living epidermis. The measurements were corrected for the differences in the refractive index of the various cutaneous layers, and the undulation of the dermo-epidermal junction. Furthermore, nucleus size and number could be assessed from horizontal optical sections. To illustrate the sensitivity of the thickness measurements, changes in the thickness of the epidermis were recorded during and after stripping of the horny layers. This non-invasive methodology is a very promising tool for morphometric studies of the living human skin at the cellular level.

Epidermal Cells↗
PubMed1990

Morphometry of endoneurial capillaries in diabetic sensory and autonomic neuropathy.

Nerve biopsies were obtained from 27 patients with diabetic neuropathy. All had a symmetric distal sensory and autonomic neuropathy or a purely sensory neuropathy. Mean age was 39.8 years (range 23-57 years). Two patients had Type 2 (non-insulin-dependent) diabetes mellitus and the remainder Type 1 (insulin-dependent) diabetes. Morphometric observations on endoneurial capillaries were compared with results from organ donor control cases and from patients with type 1 hereditary motor and sensory neuropathy. The area of the lumen of the capillaries did not differ between the three groups. The area occupied by the capillary endothelial cells in transverse section and the number of endothelial cell nuclei were increased both in the patients with diabetic neuropathy and hereditary motor and sensory neuropathy, as was the thickness of the surrounding basal laminal zone. 'Closure' of endoneurial capillaries in diabetic neuropathy, reported in another study, was not confirmed. Capillary density and nearest-neighbour distances were similar in the diabetic and organ donor control cases. Capillary density was reduced in the patients with hereditary motor and sensory neuropathy, this being related to increased fascicular area consequent upon the presence of hypertrophic changes. The presence of thickening of the pericapillary basal laminal zone and endothelial cell hyperplasia both in diabetic and hereditary motor and sensory neuropathy, the latter being a neuropathy in which a vascular basis can be discounted, makes it difficult to use such changes as an argument favouring a vascular cause for diabetic neuropathy. There were differences in the basal laminal zone between the diabetic and hereditary motor and sensory neuropathy cases suggesting that the reduplicated basal lamina was more persistent in the diabetic patients.

Adult↗
PubMed1992

The reproducibility and sensitivity of sural nerve morphometry in the assessment of diabetic peripheral polyneuropathy.

The nerve fibre loss, atrophy and injury of diabetic peripheral polyneuropathy and their responses to metabolic intervention have been studied by morphometric analysis of sural nerve biopsies. The magnitudes and sources of intra- and inter-individual variation in these morphometric measures have not been investigated previously in a systematic manner. Morphometric parameters of nerve fibre damage were measured in four separate fascicles from bilateral sural nerve specimens obtained post-mortem from 13 diabetic and 13 non-diabetic subjects. Intra- and inter-individual coefficients of variation were computed and compared to the magnitude of the differences between normal and diabetic subjects. Several morphometric variables emerged as highly sensitive and reproducible measures of nerve fibre damage suitable for clinical studies of diabetic peripheral polyneuropathy. These observations provide a rational basis for the design of future clinical trials employing morphometric end-points.

Autopsy↗
PubMed1986

Ultrastructural morphometry of epidermal Langerhans' cells: introduction of a simple method for a comprehensive quantitative analysis of the cells.

The ultrastructural demonstration of Birbeck granules is the most exact morphological marker for Langerhans' cells (LCs). We have developed a specific ultrastructural morphometric technique that enables quantitative evaluation of the numerical and volume density of LCs as well as quantification of cellular details. Evaluation of normal human skin of the hand and lower buttock showed that the epidermis of the hand contained a significantly higher numerical and volume density of LCs. At the cellular level, these LCs exhibited signs of increased metabolic activity, as reflected by the morphometric data of the nucleus, mitochondria, Golgi area, and Birbeck granules. The present technique thus allows functional characterization of LCs at the morphological level.

Adult↗
PubMed1982

Morphometry and cytodiagnosis of breast lesions.

Cells aspirated from breast lesions and smeared on slides were subjected to computer assisted morphometric analysis. Three groups of cases were studied. The first were those collected from patients with known benign and malignant lesions. The second group were 143 unselected consecutive aspirates from breast lesions and the third, a group with a needle aspirate cytodiagnosis "suspicious of malignancy". The analysis showed the malignant cells to have larger nuclei with more anisokaryosis and more variation in the nuclear cytoplasmic ratios when compared with benign cells. When this form of semiautomatic analysis was applied to the "suspicious" group the accuracy of cytodiagnosis was improved.

Biopsy, Needle↗
PubMed1980

Morphometry of intrarenal arteries in progressive sclerosis.

Vessels of known position in the vascular tree of the kidneys of two cases with a long history of progressive systemic sclerosis--one normotensive, one hypertensive--were examined morphometrically. Medial thickness, intimal thickness and the relative content of collagen and elastin in the vascular media were measured. Smooth muscle nuclei were counted in the arterial cross section. These morphometric data were compared with those obtained from two autopsy cases--one with a history of essential hypertension, one without any hypertensive history. The findings suggest that progressive sclerosis induces intimal thickening in all branches of the renal artery down to a distented diameter of 200 microns. In the case where progressive sclerosis was complicated by arterial hypertension increased medial thicknesses were found, similar to the findings in the case with a history of essential hypertension.

Arteries↗
PubMed1978

Kinetic parameters of calcium metabolism and femur morphometry in rats. I. Influence of sex and age.

The aim of this study was to establish the influence of age and sex on the kinetic and morphometric parameters of calcium metabolism. Normal values of kinetic parameters were determined in normal adult rats of different ages (4--20 months) and both sexes. Urinary hydrophyline excretion was also assessed and morphometric measurements of the femur were performed. Kinetic parameters expressed as rate constants showed a lowering of calcium turnover (kr) in the rapidly exchangeable calcium compartment until the age of 12 months and urinary and fecal rate constants (ku and kf) showed a small increase. Males exhibited a lower calcium turnover (kT) as well as lower urinary and fecal rate constants. Hydroxyproline excretion diminished with age in both sexes. Lower values were recorded in males. Morphometric parameters of femurs showed no bone loss within the examined age. The results show that differences between the sexes exist in the sense of a faster calcium turnover, higher calcium excretion rate constants, higher collagen degradation and higher bone density in adult female rats.

Aging↗
PubMed1980

Staining PO2 measurement sites in the rat brain cortex and quantitative morphometry of the surrounding capillaries.

The exact positions of microelectrodes used to measure the PO2 in the cerebral cortex of the rat were determined by staining the tissue with Alcian Blue. The measurement sites were subsequently located under a light microscope and correlated with the capillary and cellular arrangement of the cortex. The microelectrodes used for the PO2 measurements were made of gold glass fibers; the Alcian Blue was injected hydrostatically through a micropipette attached to the PO2 microelectrode. The sites where dye had been deposited were seen under a light microscope as green blue spots about 100 micrometers in diameter. The capillaries were visualized by silver nitrate perfusion. Differences between the local PO2 values in the neo- and the archeocortex were found. In the neocortex the mean PO2 was 31 mm Hg, capillary volume 1.6%, capillary surface area 980/mm2, capillary length 13.5/mm; whereas in the archeocortex these values where 21 mm Hg, 0.9%, 820/mm2 and 9.4/mm respectively. These data indicate a relationship between the microcirculatory transport system and the local oxygen tension and provide further evidence that the mean PO2 level tends to decrease when moving from the surface into the archeocortex.

Alcian Blue↗
PubMed1988

Skeletal muscle pathology in ovine congenital progressive muscular dystrophy. 2. Myofiber morphometry.

Fiber-type proportions were determined in 12 skeletal muscles (peronaeus tertius, tibialis cranialis, tensor fascia lata, psoas major, extensor digitorum lateralis, tensor fascia antibrachii, vastus intermedius, soleus, anconaeus, and flexor digitorum superficialis of fore and hind limb) from infantile (20 weeks), juvenile (16 months) and adult (2.5 and 3.5 years) Merino sheep with ovine congenital progressive muscular dystrophy and in age-matched controls. Although confined to type I fibers, lesions were severe not only in type I fiber dominant muscles (vastus intermedius, soleus and anconaeus) but also in the type II dominant medial triceps brachii and in the superficial digital flexors which had approximately equal proportions of both fiber types. The frequency distribution curves of myofiber diameter in dystrophic anconaeus and superficial digital flexor of the fore limb altered as the disease progressed. In young sheep (20 weeks) type I fibers increased in size, then in juvenile and adult sheep the curves became flatter and broader and lost symmetry due to the presence of hypertrophic and atrophic fibers. The fiber diameter profiles were similar to those seen in Duchenne dystrophy in humans.

Animals↗
PubMed1977

Splanchnic preganglionic neurons in man. I. Morphometry of preganglionic cytons.

The thoracic sympathetic preganglionic outflow is important in the maintenance of postural normotension in man. Normative data on the intermediolateral column (ILC) neuron cell bodies is lacking. Counts and measurements on the right ILC cytons have been performed on the T6, T7 and T8 segments of 12 spinal cords of man obtained within 4-6 h of death. A methodology was employed which has been optimized to prevent shrinkage of cytoplasm from the neuropil and which mathematically corrects the number of cytons of neurons and their frequency distribution for split cell error. The mean cyton counts for T6, T7 and T8 spinal cord segments were 5002, 5004 and 4654 respectively. No significant sex difference was shown. Most cytons ranged in diameter from 8-23 micrometer; the major peak in the size histogram is at 12-13 micrometer and a smaller peak is seen at 16 micrometer. There is a progressive reduction of ILC cytons with age. In adult man approximately 370 cytons (approximately 8%) are lost per decade. This reduction in ILC cytons with age may be a morphological basis for postural hypotension of the aged.

Adult↗
PubMed1980

Morphometry of astrocyte and oligodendrocyte ultrastructure after portocaval anastomosis in the rat.

The ultrastructure of astrocytes and oligodendrocytes was investigated in rats 10 days, 30 days, and 10 weeks after portocaval anastomosis (PCA). Cell and nuclear sizes were measured by planimetry on randomly sampled cells magnified X 24,000. The volume fractions of mitochondria, glia fibrils, and lipofuscin granules were measured in astrocytes by electronic image analysis. The mitochondrial profile area distribution and oligodendrocyte mitochondrial content were likewise estimated. All PCA animals had an increased astrocyte cell and cytoplasmic area, and after correction for cytoplasmic edema all groups had an enhanced mitochondrial fraction and mitochondrial number. The mitochondrial sizes were increased in all PCA groups. The mitochondrial profile area distribution curves did not suggest more than one group of mitochondria. All PCA groups had increased fractions of lipofuscin granules and glia fibrils. The oligodendrocytes had a slight fall in cell, nuclear, and cytoplasmic area after 30 days of shunting, and the mitochondrial fraction was diminished. After 10 weeks of PCA, all changes were reversed to normal values. It is concluded that the astrocytes are the active cells in the brain metabolism of ammonium. The oligodendrocytes seem to be dependent on neuronal integrity and do not contribute to the brain ammonium metabolism. The increase in astrocyte lipofuscin granules content may be explained by a beginning neuronal loss.

Animals↗
PubMed1987

Morphometry of the degenerative process in the hypoglossal nerves in amyotrophic lateral sclerosis.

The peripheral hypoglossal nerves in 13 cases of amyotrophic lateral sclerosis (ALS) and five control cases were examined using morphometrical methods to demonstrate the degenerative process of motor nerve degeneration. The total number of myelinated fibers and their histograms were analyzed according to the degree of severity of the degeneration. Reduction of the total number of myelinated fibers in ALS hypoglossal nerves were graded in three groups: mild 65%-75%, moderate 50%-65% and severe 30%-50% of the myelinated fibers in controls. Each histogram of the remaining myelinated fibers showed different patterns corresponding to the degree of the degeneration and disclosed that the progressive reduction of large myelinated fibers was the fundamental change. Small myelinated fibers were not reduced, but increased, especially in the group with a moderate grade of degeneration. In plastic section, there were clusters of regenerated myelinated fibers. The transient increase of small myelinated fibers may be a reflexion of myelinated fiber regeneration during the progressive degenerative process of the motor neurons. The correlation between the degree of severity of the hypoglossal nerve degeneration and the atrophy of the tongue muscle and the duration of bulbar symptoms was examined and discussed.

Amyotrophic Lateral Sclerosis↗