Stereological Evaluation Research Articles

Stereological evaluation and Golgi study of the sexual dimorphisms in the volume, cell numbers, and cell size in the medial preoptic nucleus of the rat.

The medial preoptic nucleus (MPN) and the sexually dimorphic nucleus of the preoptic area (SDN-POA) stand out as prominent sexually dimorphic cell groups of the rat brain. However, quantitative data on sex-related differences in these nuclei in the adult rat are confined to their volume. We have used stereological methods and Golgi-impregnated material to examine whether, in young adult rats, the sexual dimorphism in the volume of the MPN, including its divisions, and of the SDN-POA, reflect similar differences in the number and size of their neurons. We found that the total number of neurons in all MPN divisions is higher and the mean somatic volume larger in males than in females. In addition, the total dendritic length of MPN neurons is greater, but the dendritic spine density is smaller, in males than in females. Likewise, in the SDN-POA the total number and size of its neurons is greater in males than in females. The sex differences in all quantitative parameters evaluated accounted for the larger volume of the MPN and SDN-POA in males relative to females. In addition, the MPN neuropil also displays sex-related differences in its volume, and these differences closely match those detected for the volume of each MPN division. It deserves to be emphasised that the numerical density of neurons was the only parameter found to be significantly higher in females than in males in all MPN divisions and in the SDN-POA. Our results show that the MPN and the SDN-POA display sex differences in the volume, total number of neurons, and size of neuronal cell bodies and dendritic trees. Furthermore, they also indicate that the neuropil is critical for the establishment of sexual dimorphism in the size of the MPN.

Volumetric determination of normal and scoliotic vertebral bodies.

A new method is presented for stereological evaluation of the volume of the vertebral body in vivo. The height of the vertebral body is measured at three standardised points on an anteroposterior radiograph and at two other points on a lateral one. The area of the body is also measured using a special grid superimposed on a CT scan from the middle part of the vertebra. The volume of the vertebral body is then calculated using Cavalieri's principle for irregular objects: V = delta a x H, where V is the volume of the vertebral body, delta a is the mean cross-section surface area on the CT scan and H is the mean of the heights at the five points on the radiographs, computed as mean weighted circumferential height. The volume of one normal and one scoliotic vertebra was evaluated in vitro using this formula. The obtained values were compared with the values derived from serial CT scans of the two vertebrae. The results showed that the volume of the normal vertebra measured with our new method was 15.9 cm3 and measured with serial CT scans using the same grid it was 15.07 cm3. For the scoliotic vertebra the values were 17.6 and 17.3 cm3, respectively. The degree of accuracy of the measurements with the presented method as compared with the serial CT method was 95% for the normal and 98.5% for the scoliotic vertebra. To prove the clinical applicability of the method, the heights of the apical and of the upper and the lower end vertebrae of the curve and the volume of the apical vertebrae were evaluated in eight scoliotic girls (nine curves) before and 3 years after spinal instrumentation and posterior fusion. The results showed that the mean circumferential height of the three vertebrae had increased significantly at the last follow-up. The volume of the apical vertebra had also increased, but the difference was not significant. It is concluded that the described method is easy to apply and has satisfactory accuracy for in vivo longitudinal studies of the volume of the vertebral body on radiographs and CT scans.

Stereological Evaluation of Human Spermatogenesis after Suppression by Testosterone Treatment: Heterogeneous Pattern of Spermatogenic Impairment

Stereological Evaluation of Human Spermatogenesis after Suppression by Testosterone Treatment: Heterogeneous Pattern of Spermatogenic Impairment

Stereological evaluation of human spermatogenesis after suppression by testosterone treatment: heterogeneous pattern of spermatogenic impairment.

Testosterone (T) treatment suppresses gonadotropin levels in normal men and is a promising reversible contraceptive that induces azoospermia in approximately 70% of subjects and oligospermia in the remainder; however, the basis of this variable response is unclear. This study aimed to investigate this reported variable response by examining the spermatogenic process and quantitating germ cell number in men after T-induced gonadotropin withdrawal. Ten normal fertile men (31-46 yr), already planning to undergo vasectomy, either received T enanthate (200 mg, i.m., weekly) for 19-24 weeks (n = 5; TE group) or proceeded directly to surgery (n = 5; controls), at which time a unilateral testicular biopsy was taken, and germ cell numbers were estimated using the optical disector stereological method. In response to TE treatment, serum T levels rose 2-fold, and FSH/LH levels became undetectable. Sperm counts fell to azoospermia in 4 men and to 21 million/mL in the fifth man. The mean number of type A spermatogonia per 100 Sertoli cells was unchanged, but type B spermatogonia fell markedly to 10% of the control values, and later germ cell types decreased to 11-18% of the control values. The pattern of germ cell suppression varied widely and showed no relationship with sperm count or the time to azoospermia. Despite the presence of elongated spermatids (1.4-20% of the control), four men remained azoospermic. Two TE subjects with similar early germ cell complements and elongated spermatid numbers had sperm counts of zero and 21 million/mL; the latter man demonstrated marked variability in germ cell numbers between adjacent tubules. We conclude that 1) the principal spermatogenic lesion in TE-treated men is the marked (90%) inhibition of type A-->B spermatogonial maturation. Other sites are also affected, particularly the release and/or survival of elongated spermatids during transit; and 2) a steady state in germ cell number may not be established even after 4-5 months of TE treatment. The findings suggest that TE treatment does not adequately or consistently withdraw hormonal support for spermatogenesis, leading to variable between- and within-individual patterns of germ cell suppression.

Recent Developments in Automated Crystal Orientation Mapping (ACOM) - Quantitative Evaluation and Graphical Representation of Individual Grain Orientation Data

A system is presented which performs fully unattendedly the measurement of the crystal orientations of individual grains using the automated interpretation of Backscatter Kikuchi Patterns (BKP). The features which are necessary to scan a selected area of the specimen surface with a high spatial resolution are described Several possibilities for a quantitative evaluating of the acquired orientation data and their representation by pseudo-colored maps are shown. A simple estimate of the quality of BKD pattern yields information on the degree of deformation or recrystallization. Methods of quantitative stereological and statistical evaluation of the acquired data are demonstrated. The local texture of small selected areas can be investigated and be described by its statistical functions which are calculated from the achieved orientation data.

Neonatal administration of FSH increases Sertoli cell numbers and spermatogenesis in gonadotropin-deficient (hpg) mice.

We previously demonstrated that androgens alone, in the complete absence of gonadotropins, initiated qualitatively complete spermatogenesis in hypogonadal (hpg) mice. Although germ cell to Sertoli cell ratios were normal in hpg mice with androgen-induced spermatogenesis, testicular size. Sertoli cell and germ cell numbers only reached 40% of those of non-hpg mice, and Sertoli cell numbers were unaffected by androgen treatment started at 21 days of age. We postulated that these observations were due to diminished gonadotropin-dependent. Sertoli cell proliferation during perinatal life while the Sertoli cells still exhibited normal carrying capacity for mature germ cells. In order to test this hypothesis, we examined the effects of administering androgens and gonadotropins to hpg mice during the first 2 weeks of postnatal life when Sertoli cells normally continue to proliferate. The study end-points were Sertoli and germ cell numbers in hpg mice following induction of spermatogenesis by 8 weeks treatment with 1 cm subdermal silastic testosterone implants. Newborn pups (postnatal day 0-1) were injected s.c. with recombinant human FSH (rhFSH) (0.5 IU/20 microliters) or saline once daily for 14 days, with or without a single dose of testosterone propionate (TP) (100 micrograms/20 microliters arachis oil) or human chorionic gonadotropin (hCG) (1 IU/20 microliters). Untreated hpg and phenotypically normal littermates were studied as concurrent controls. At 21 days of age, all treated weaning mice received a 1 cm silastic subdermal testosterone implant and, finally, 8 weeks after testosterone implantation, all mice were killed. As expected, qualitatively complete spermatogenesis was induced in all groups by testosterone despite undetectable circulating FSH levels. Exogenous rhFSH increased testis size by 43% (P < 0.002) but a single neonatal dose of either TP or hCG reduced the FSH effect although neither TP nor hCG had any effect alone. In contrast, a single neonatal dose of TP or hCG increased final seminal vesicle size whereas FSH had no effect. FSH and TP treatment significantly increased absolute numbers of testicular spermatids compared with saline treatment, whereas hCG and TP significantly increased testicular sperm when expressed relative to testis size. Stereological evaluation of Sertoli and germ cell numbers demonstrated a rise in the absolute numbers of Sertoli and all germ cell populations induced by neonatal administration of hormones. When expressed per Sertoli cells the numbers of germ cells in the treated mice were between 85 and 90% of non-hpg controls. We conclude that exogenous FSH treatment during the first 2 weeks of postnatal life, coinciding with the natural time of Sertoli cell proliferation, increases Sertoli cell numbers and thereby the ultimate size of the mature testis and its germ cell production. Thus neonatal gonadotropin secretion may be a critical determinant of the sperm-producing capacity of the mature testis. In addition, neonatal exposure to androgens could be important for the imprinting of sex accessory organs in hpg mice, with the long-term effects of altering the sensitivity of the accessory organs to exogenous testosterone later in life.

Infection with Trypanosoma brucei potentiates the antifertility effect of gossypol, especially in the protein-malnourished male rat.

Reproductive parameters were investigated in gossypol-treated male rats that had been infected experimentally with Trypanosoma brucei and then place on one of two diets of differing (low and normal) protein content. The endpoints assessed were reproductive organ weights, semen epididymal sperm counts, serum testosterone and histological, stereological and histomorphometric evaluation of the testis and accessory reproductive organs. Most of the parameters studied were lowest in the protein-malnourished, gossypol-treated, trypanosome-infected animals, when compared to those obtained from the corresponding animals that were only gossypol-treated or trypanosome-infected. Mean testicular size and the diameter and the total length of seminiferous tubules were especially reduced, indicating that the overall volume of the seminiferous epithelium in these animals was smaller. These findings suggest that reproductive capacity could be impaired in protein-malnourished, trypanosome-infected animals fed on gossypol-containing products, even when there are no obvious clinical signs of disease. This could translate into increased production costs in a farm enterprise. Screening for haemoparasitism would also seem to be worthwhile prior to evaluation and/or use of gossypol (or perhaps other potential contraceptives) in human subjects, especially in communities that are confronted with severe food shortages.

Stereological evaluation of vascular adaptations in human placental villi to differing forms of hypoxic stress

The aim was to determine whether the fetal vasculature of the human placenta adapts in a uniform manner to different forms of hypoxic stress. Stereological analyses were performed on the intermediate and terminal villi of placentae obtained from each of the following conditions; high altitude (up to 2800 m), maternal iron-deficiency anaemia and pre-eclampsia. These conditions were taken to represent hypoxic, anaemic and ischaemic hypoxia, respectively. In each situation, there was a trend for the volume fraction of the fetal capillaries to be higher than in the controls, although the differences were statistically significant only for the cases of hypoxic and anaemia hypoxia. This was due, in part, to capillaries being of larger diameter. Evidence for capillary proliferation was inconsistent. At high altitude there was a trend for the capillary length density to be raised, but there was no change in the ratio of capillary:villous length. In both maternal anaemia and pre-eclampsia the capillary length density remained constant, but the ratio of capillary:villous length was significantly raised in the former. These results suggest that dilatation of the capillary sinusoids, with accompanying thinning of the villous membrane, is the principal adaptation to hypoxia under the conditions studied.

A Comparison Between the Effects of Paraffin and Plastic Embedding of the Normal and Obstructed Minipig Detrusor Muscle Using the Optical Dissector

Purpose The purpose of this study was to estimate the relative shrinkage of the normal, obstructed and recovery minipig urinary bladder by comparing tissues from the same bladders embedded in paraffin or plastic. Materials and Methods Optical dissectors were used to make nucleus numerical density estimates in thick paraffin and plastic sections from the same bladder. The ratio of the 2 numerical densities depends only on differences in tissue shrinkage. In 9 minipigs a partial bladder outlet obstruction was created by implanting a 6 to 7 mm. Teflon ring around the proximal urethra. After an obstruction period (median 63 days) the ring was removed and after a recovery period (median 60 days) the animals were sacrificed. Biopsis were taken prior to obstruction, at removal of obstruction, and after recovery and were processed for paraffin and plastic sections to evaluate relative shrinkage. Two control pigs were sham-operated and biopsies taken at the same 3 time points. Results The optical dissector method was found to be an easy way to estimate the relative shrinkage of paraffin-embedded bladder tissue in proportion to plastic-embedded tissue. Both in human and minipig bladders, paraffin embedding caused a relative shrinkage of about 30 percent in proportion to plastic embedding. Both the obstructed and recovery detrusor muscles responded to embedding by either method in a manner indistinguishable from the normal bladder. Conclusion When dealing with stereological evaluation of the detrusor muscle, the relative shrinkage of the embedded normal, obstructed and recovery bladder tissue can be ignored.

Morphometric study of focal cerebral ischemia in rats: a stereological evaluation

An efficient, unbiased stereological method for estimating and evaluating volumetric parameters of cortical infarction is presented. Long-Evans rats were subjected to a standard procedure for achieving focal cerebral ischemia, consisting of permanent ligation of the right middle cerebral artery and temporary occlusion of both common carotid arteries. Animals were killed 24 h later by perfusion fixation and the brains were embedded in celloidin, serially sectioned at 60 μm and stained with Cresyl violet. All cases showed identifiable areas of infarction of variable extent within the territory of the right middle cerebral artery. Involvement of allocortical or subcortical structures was variable and in three cases there were small foci of infarct in the contralateral hemisphere. The absolute volumes of the infarcted tissue and of the spared cortex of both hemispheres were obtained by means of an unbiased estimator of volume based on Cavalieri's principle. The best estimate of the actual amount of infarcted cortex was found to be given by the volume ratio between the spared cortex of the right (infarcted) hemisphere and the total cortex of the left (control) hemisphere. This approach avoided the error introduced by the accompanying edema and decreased the observed interanimal variance. Sample size predictions for therapeutic-pharmacological trials show that relatively few animals would be needed using the model of ischemia and quantitative morphometry presented here to detect a 30–40% change in infarct volume.

Molecular biology of human male infertility: links with aging, mitochondrial genetics, and oxidative stress?

Molecular biology of human male infertility: links with aging, mitochondrial genetics, and oxidative stress?

Subcellular volumes and metabolite concentrations in barley leaves

Metabolite concentrations in subcellular compartments from mature barley (Hordeum vulgare L. cv. Apex) leaves after 9 h of illumination and 5 h of darkness were determined by nonaqueous fractionation and by the stereological evaluation of cellular and subcellular volumes from light and electron micrographs. Twenty one-day-old primary leaves of barley with a total leaf volume of 902 μL per mg chlorophyll were found to be composed of 27% epidermis, 42% mesophyll cells, 6% veins, 4.5% apoplast and 23% gas space. While in epidermal cells 99% of the volume was occupied by the vacuole, mesophyll cells with an average volume of 31.3 pL consisted of 23 pL (73%) vacuole, 4.6 pL (19%) chloroplasts, 2.06 pL (6,7%) cytosol (including smaller organelles and vesicles), 0.34 pL (1%) mitochondria and 107 fL (0.34%) nucleus. The differences between leaves harvested after 9 h of illumination and after 5 h of darkness were in the size of the stromal compartment and the starch grains therein. Subcellular metabolite concentrations were calculated from the compartmental volumes and metabolite contents of the compartments as determined by nonaqueous fractionation. The amino-acid concentrations in stroma and cytosol were rather similar after 9 h of illumination and 5 h of darkness. In contrast, the vacuolar amino-acid concentrations were about one order of magnitude lower than the stroma and cytosol values, and there was a slight increase in concentration after 5 h of darkness.

A T.E.M. stereological analysis of blood vessels and nerves in marmoset periodontal ligament following endodontics and magnetic incisor extrusion.

Maxillary incisor fracture is a common dental injury. It is frequently treated by endodontic therapy, orthodontic extrusion, and a retention period. The aim of the study was to mimic this regime and stereologically quantify the vascular and neural reconstitution of the periodontal ligament (PDL). Ten marmosets (Callithrix jacchus) had the right maxillary incisor crown removed, root canal treatment, and magnetic incisor root extrusion of 1.2 mm. After incisor retention for 9 weeks, the animals were perfusion fixed for electron microscopy. The incisor PDL was sectioned at 250 micron intervals from the cervical to apical region and recorded for stereological evaluation. Further evaluations were conducted with a two-way analysis of covariance using program 3V of the BMDP-81 statistical software package. The total luminal PDL vascular volume was statistically unchanged. An increased occurred in the mean number of venous capillaries from 0.07 +/- 0.3 to 0.17 +/- 0.3 per 560 microns 2 of PDL (P < 0.05). Vessel wall and luminal volume of post-capillary-sized venules decreased from 1.64 to 1.03 per cent, and 5.32 to 3.04 per cent (P < 0.05), respectively. Mean luminal and abluminal diameters for post-capillary-sized venules were reduced (P < 0.05). Myelinated and unmyelinated axons showed no significant change in volume (Vv), length (Lv), or surface (Sv) densities. Axon number/unit, PDL area, and mean diameters demonstrated no statistical differences. Since the PDL demonstrated almost complete morphological re-establishment of the microvascular and axon systems, it is considered that this clinical treatment is a justifiable rationale.

Stereological evaluation of altered hepatocellular foci in control Wistar rats.

Quantitative evaluation and stereological analysis of altered hepatocellular foci (AHF) were performed on hematoxylin and eosin-stained (H&E) liver sections from control Wistar rats from 9 2-yr carcinogenicity studies conducted between 1981 and 1991. Morphologic criteria previously described were used to classify AHF in H&E stained livers into basophilic, eosinophilic, clear, mixed, or vacuolated cell foci. Hepatocellular adenomas were seen in 1.6% of control rats and carcinomas were seen in 0.2%. AHF were diagnosed in 20% of control Wistar rats and were seen only in rats dying after 47 weeks. Basophilic foci were the most common AHF seen; incidences of tigroid and diffuse basophilic AHF were similar. Vacuolated foci were not identified in any rat. The mean number of AHF per cubic centimeter of liver was 101. The mean AHF volume varied from 0.002-0.048 mm3 and the mean volume fraction ranged from 0.005-0.026%. Compared to AHF in aged control Fischer 344 rats, AHF in control Wistar rats were fewer and smaller.

Ultrastructural morphometric analysis of the uterine epithelium during early pregnancy in the sheep

Stereological techniques were used to quantify ultrastructural changes in the caruncular epithelium during the pre- (Day 13), peri- (Day 16) and post- (Days 19 and 22) attachment periods of placentation. Tissues from Day-13 non-pregnant ewes were used as controls. Uteri for stereological evaluation were perfused via the uterine artery with 3% glutaraldehyde and separated into proximal, middle and distal regions. Tissues from caruncular areas were processed for electron microscopy. Volume fractions (Vv) of nuclei, mitochondria, lipid and cytoplasmic granules were estimated by point-counting volumetry. Surface areas per unit tissue volume (Sv) of mitochondrial membranes and cristae, Golgi, plasmalemma, endoplasmic reticulum and nuclear membranes were estimated by line-intersection counting. The only significant difference between pregnant and non-pregnant uterine epithelium at Day 13, a time before attachment, was a lower Sv of smooth endoplasmic reticulum (SER) in tissue from pregnant ewes. This value returned to control (non-pregnant Day 13) levels at Day 16, and was again significantly reduced at Days 19 and 22. The Vv of lipid decreased significantly at Day 16 and remained at reduced levels thereafter. These changes may reflect the effects of conceptus products on lipid storage and mobilization. The Sv of rough endoplasmic reticulum (RER) significantly increased on Day 16 of gestation, and remained elevated on Day 19. These results may reflect increased synthesis of protein for export at these times. In general, several of the values measured which may be indicative of cellular metabolism were reduced at Day 22 of pregnancy, perhaps suggesting diminished metabolism by the uterine epithelium after attachment of the trophoblast.(ABSTRACT TRUNCATED AT 250 WORDS)

Origin of noradrenergic innervation of the spleen in rats

Noradrenergic (NA) sympathetic innervation of the spleen was examined in young adult Sprague-Dawley rats (200–250 g) following surgical removal of the superior mesenteric-celiac ganglia (SM-CG) and/or bilateral transection of the subdiaphragmatic vagus nerve. Sham-operated and unoperated rats served as controls. NA sympathetic innervation of spleens from sham-operated and unoperated controls, and from vagotomized rats, was qualitatively similar, with fibers distributing to the capsule, trabeculae, vasculature, and parenchyma of the white pulp. Complete ganglionic extirpation resulted in almost total denervation of NA fibers in all compartments of the spleen. High-performance liquid chromatography with electrochemical detection (LCEC) for catecholamines (CA) and quantitative morphometry of the density of NA varicosities confirmed these observations. LCEC revealed a greater than 85% depletion of norepinephrine (NE) in the spleen following superior mesenteric-celiac ganglionectomy. Stereological evaluation of NA varicosities with a point counting method revealed a decline of 99% in the volume density of NA terminals that occurred uniformly in all compartments of spleens from ganglionectomized rats. In addition, stereological analysis revealed a loss of total NA varicosities (approximately 31% decrease) in spleens from sham-operated rats. This loss in volume density occurred largely due to a loss in parenchymal fibers (approximately 45% decrease). Bilateral subdiaphragmatic vagotomy blocked the effect on NA innervation produced by the surgical stress of sham operation. Retrograde tracing following injection of either fluorogold or true blue into the spleen, coupled with immunocytochemical localization of tyrosine hydroxylase (TH), demonstrated abundant fluorogold (true blue)-labeled neurons in the SM-CG; many, but not all, of these neurons also were TH-positive. These findings indicate that the SM-CG neurons supply NA innervation to the spleen, providing sympathetic innervation as the second neuron in the classical two-neuron sympathetic chain, and suggest additional non-NA innervation of the spleen as well. This study also suggests that surgical stress of sham operation may alter directly the NA innervation of the spleen, possibly by inducing temporary retraction of NA fibers of the parenchymal compartment, which is likely to reduce the availability of NE for interaction with cells of the immune system that possess adrenoceptors and are present adjacent to NA varicosities in this region. Bilateral vagotomy ameliorated the effects of sham operation on NA innervation; since the vagal nerve does not distribute fibers to the spleen, this effect is likely to occur through altered feedback circuits effecting sympathetic outflow, or through altered neuroendocrine outflow.

Proliferative activity of cutaneous melanocytic tumors defined by Ki-67 monoclonal antibody. A quantitative immunohistochemical study.

In various tumors, proliferative activity has been demonstrated to be closely related to the degree of malignancy. In the present study, we examined the proliferative pool in a total of 25 melanocytic (nevocytic) skin tumors by immunohistochemical staining with Ki 67 monoclonal antibody (labeling the G1, S, G2-phase of the cell cycle) and quantitative morphological evaluation. There were highly significant differences in the numerical density of Ki 67-positive cells between nevocellular nevi (number of positive cells: 2.2 +/- 0.7 X 10(3)/mm3), primary malignant melanoma (6.3 +/- 1.9 X 10(3)/mm3), and metastatic malignant melanoma (47.0 +/- 9.2 X 10(3)/mm3). Within the malignant melanoma group, there was a significant correlation between proliferative activity and maximum tumor thickness (r = 0.85; p less than 0.05). Comparison with previous studies using tritiated thymidine labeling and colony-forming efficiency shows that the stereological evaluation of Ki 67 staining in melanocytic skin tumors provides a reliable estimate of proliferative activity, which might be of prognostic value.

Quantitative stereological evaluation of four histochemical markers of altered foci in multistage hepatocarcinogenesis in the rat.

Female F344/N rats dosed with diethylnitrosamine (DEN) 24 h after partial hepatectomy were treated with the promoting agents, phenobarbital (PB) or 3,4,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or the peroxisome proliferating agent, WY 14,643, for 6 months. Another group was subjected to the Solt-Farber protocol. Altered hepatic foci (AHF) were analyzed by quantitative stereology from frozen serial sections stained for gamma-glutamyl transferase (GGT), canalicular adenosine triphosphatase (ATPase), glucose-6-phosphatase (G6Pase) and the placental isozyme of glutathione S-transferase (PGST). PGST scored more foci in all groups than GGT and ATPase. PGST marked greater focal volume than GGT or ATPase, and PGST marked focal volume equal to or greater than G6Pase in rats treated with PB, TCDD or the Solt-Farber protocol. However, after treatment with WY 14,643, GGT and PGST marked much less focal volume than ATPase or G6Pase, and PGST scored fewer foci than G6Pase. Numerical estimations of foci scored by those markers on the basis of area of the entire tissue section (per cm2) were relatively different from those values determined by quantitative stereology. While these results confirm earlier studies, they demonstrate the importance of quantitative stereologic analysis of AHF during multistage hepatocarcinogenesis.

Stereological substructure analysis in hot-deformed metals from TEM-images

In the present paper three possible methods of stereological evaluation of particle arrangements are discussed: Practical application of the procedures is demonstrated by results which were obtained from hot-deformation of several material under different hot-working conditions. It can be shown that stereological interpretation of TEM images renders refined conclusions concerning the mechanisms of structure changes occurring in hot-deformation.

Geometric model of the rat intestinal mucosa for stereological evaluation of villus amplification factors.

This study was prompted by a desire to compare villus amplification factors at various sites along the small intestine of the laboratory rat. The paper describes a precise and efficient stereological method for achieving this objective. The method relies on conventional intersection counts performed exclusively on transverse sections. Analyses are based on six adult rats. Pieces of small bowel fixed by in situ perfusion were taken from proximal, middle and distal regions. Intersection counts on villus and primary mucosal surfaces were performed on transverse and longitudinal sections through the same regions. These counts revealed that intestinal architecture can be reasonably approximated by the simple geometric model of an isotropic villus surface on an anisotropic (cylindrical) primary mucosal surface. As a result, intersection counts may be confined to transverse sections (to reduce sampling variance) and the appropriate ratio estimator multiplied by the numerical coefficient 4/pi (to obtain an estimate of surface amplification due to villi).