Wiley
0003-276X
1097-0185
Cơ quản chủ quản: N/A
Lĩnh vực:
Các bài báo tiêu biểu
Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator Abstract A stereological method for obtaining estimates of the total number of neurons in five major subdivisions of the rat hippocampus is described. The new method, the optical fractionator, combines two recent developments in stereology: a three‐dimensional probe for counting neuronal nuclei, the optical disector, and a systematic uniform sampling scheme, the fractionator. The optical disector results in unbiased estimates of neuron number, i.e., estimates that are free of assumptions about neuron size and shape, are unaffected by lost caps and over‐projection, and approach the true number of neurons in an unlimited manner as the number of samples is increased. The fractionator involves sampling a known fraction of a structural component. In the case of neuron number, a zero dimensional quantity, it provides estimates that are unaffected by shrinkage before, during, and after processing of the tissue. Because the fractionator involves systematic sampling, it also results in highly efficient estimates. Typically only 100–200 neurons must be counted in an animal to obtain a precision that is compatible with experimental studies. The methodology is compared with those used in earlier works involving estimates of neuron number in the rat hippocampus and a number of new stereological methods that have particular relevance to the quantitative study of the structure of the nervous system are briefly described in an appendix.
Tập 231 Số 4 - Trang 482-497 - 1991
Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action Abstract The properties of the inorganic dye ruthenium red are presented with emphasis upon its use for electron microscopy of cells and tissues. Although commercial ruthenium red often can be used directly, it always contains various impurities and by‐products. One of these, termed ruthenium violet, can be isolated and is useful by itself. Absorption spectra of the ruthenium dyes and common impurities are given so that an assay is possible for any sample. Convenient fixative recipes containing ruthenium red or violet are provided together with constraints necessary for a reliable reaction to label extracellular acidic mucosubstances. Perfusion was not successful. The specificity of the ruthenium red reaction was evaluated by spot testing with 57 substances, and by titration with chemically defined pectins. The results indicate that ruthenium red, as a hexavalent cation, precipitates a large variety of polyanions by ionic interaction, and that its classical reaction with pectin is typical rather than specific. New data are presented regarding its reaction with phospholipids. For electron microscopy, a further reaction with OsO4 amplifies the feeble electron density, which is the counterpart of its intense optical labeling, to a practical level resulting in strong contrast. An hypothesis is presented for the mechanism underlying this intensification.
Tập 171 Số 3 - Trang 347-368 - 1971
Satellite cells as the source of nuclei in muscles of growing rats Abstract The source of the new nuclei appearing during the growth of muscle fibers was examined in the tibialis anterior muscle of young Sherman rats (14–17 days of age) using radioautography at various intervals after a single injection of a small, non‐toxic dose of 3 H‐thymidine (2 μCi/g body weight). Two techniques were employed: (1) labeled nuclei were detected in 1 μ thick radioautographs examined in the light microscope, and identified by simultaneous electron microscope examination of an adjacent section. The nuclei were then classified either as “true” muscle nuclei (within the plasmalemma of the fibers) or as belonging to “satellite cells” (which are mononucleated cells with scanty cytoplasm wedged between plasmalemma and basement membrane). (2) Muscle fibers freed by collagenase digestion were radioautographed one hour after 3 H‐thymidine injection in order to determine the total number of labeled nuclei (true muscle nuclei plus those of satellite cells) per unit length of fiber. Certain nuclei within the basement membrane of muscle fibers are labeled one hour after 3 H‐thymidine and, therefore, synthesize DNA. The electron microscope demonstrates that these nuclei invariably belong to satellite cells, never to true muscle nuclei. Furthermore, the total number of labeled nuclei per unit length of fiber doubles between 1 and 24 hours; and, therefore, the labeled satellite cell nuclei undergo mitosis. Following mitosis, half of the daughters of satellite cells are incorporated into the fibers to become true muscle nuclei. The remaining half divides again later; and half of their daughter cells are incorporated. Thus, satellite cells in young rats divide repeatedly and function as a source of true muscle nuclei.
Tập 170 Số 4 - Trang 421-435 - 1971
The identification, origin, and migration of the primordial germ cells in the mouse embryo
Tập 118 Số 2 - Trang 135-146 - 1954
The human cadaver in the age of biomedical informatics Abstract Major national and international critiques of the medical curriculum in the 1980s noted the following significant flaws: (1) over‐reliance on learning by rote memory, (2) insufficient exercise in analysis and synthesis/conceptualization, and (3) failure to connect the basic and clinical aspects of training. It was argued that the invention of computers and related imaging techniques called to question the traditional instruction based on the faculty‐centered didactic lecture. In the ensuing reform, which adopted case‐based, small group, problem‐based learning, time allotted to anatomical instruction was severely truncated. Many programs replaced dissection with prosections and computer‐based learning. We argue that cadaver dissection is still necessary for (1) establishing the primacy of the patient, (2) apprehension of the multidimensional body, (3) touch‐mediated perception of the cadaver/patient, (4) anatomical variability, (5) learning the basic language of medicine, (6) competence in diagnostic imaging, (7) cadaver/patient‐centered computer‐assisted learning, (8) peer group learning, (9) training for the medical specialties. Cadaver‐based anatomical education is a prerequisite of optimal training for the use of biomedical informatics. When connected to dissection, medical informatics can expedite and enhance preparation for a patient‐based medical profession. Actual dissection is equally necessary for acquisition of scientific skills and for a communicative, moral, ethical, and humanistic approach to patient care. Anat Rec (New Anat) 269:20–32, 2002. © 2002 Wiley‐Liss, Inc.
Tập 269 Số 1 - Trang 20-32 - 2002
Degeneration of germ cells in normal, hypophysectomized and hormone treated hypophysectomized rats Abstract In normal adult rats some germ cells degenerate at several vulnerable steps of spermatogenesis. These are the type A spermatogonia, midpachytene spermatocytes, primary and secondary spermatocytes which degenerate during their respective maturation divisions and step 7 and 19 spermatids. In the present study, these degenerating cells were examined under the electron microscope, and their frequency was determined in toluidine blue stained semithin sections of testes from normal, hypophysectomized (at 5.5 days after operation) and hypophysectomized rats injected with FSH and LH separately or in combination. With the exception of the step 19 spermatids, the degenerating germ cells underwent necrosis in vacuolated spaces delimited by Sertoli cells. In the case of the affected step 19 spermatids, an apical cytoplasmic process of the Sertoli cell initially ensheathed a long segment of their flagellum, and then each degenerating cell was drawn deep in the seminiferous epithelium where it was phagocytozed by the Sertoli cell. Soon after hypophysectomy the incidence of degenerating mid‐pachytene spermatocytes, step 7 and 19 spermatids which are present in stages VII or VIII of the cycle of the seminiferous epithelium, increased significantly. In contrast the number of degenerating primary or secondary spermatocytes during the meiotic divisions seen in stage XIV of the cycle or of any other germinal cell was not significantly modified. While the injection of FSH alone had no influence on the number of degenerating cells in hypophysectomized rats, injections of LH at the two doses administered (0.7 μg or 20 μg) reduced significantly the number of degenerating cells seen in stages VII‐VIII of the cycle; combined injections of FSH and LH (20 μg) reduced the number of these degenerating cells to the normal low values. Thus it appeared that the mid‐pachytene spermatocytes and the step 7 and 19 spermatids, all present in the adluminal compartment of the seminiferous epithelium in stages VII or VIII of the cycle, were more sensitive to the presence of absence of gonadotropic hormones than the other germ cells present in the seminiferous epithelium.
Tập 187 Số 3 - Trang 347-365 - 1977
Composition of trabecular and cortical bone Abstract A technique was devised for obtaining marrow‐free trabecular bone so that the trabecular and cortical bone composition of dog, steer, monkey and man could be studied. Vertebral body was cut in the frontal plane into slices 2 mm thick with an electric band saw. The cortical bone encrircling each slice was trimmed off with a cleaver, leaving intact a lacy slice of trabecular bone. The marrow substance was washed away by fine and powerful jets of tap water obtained from an adjustable nozzle. Marrow washing was discontinued when no trace of red color was visible on holding the slice in front of a strong light source. The quantities of water, volatile inorganic, organic and ash fractions in cortical bone was alike for all species. Likewise, the trabecular bone fractions of each was quantitatively similar. In general, density and amount of ash in cortical bone was higher than that in trabecular bone. The water and ash:organic ratio were higher in trabecular than in cortical bones. Ratios of trabecular to cortical ash for the bones in an entire dog skeleton were also determined. Based on this, volume or mass of the various bone fraction may be estimated when the weight of the ash for any particular bone is known.
Tập 149 Số 3 - Trang 325-331 - 1964
Scanning electron microscope study of the developing microvasculature in the postnatal rat lung Abstract During postnatal growth the parenchymal septa of rat lung undergo an impressive restructuring. While immature septa are thick and contain two capillary layers, mature septa are slender and contain a single microvascular network. Using the Mercox® casting technique and scanning electron microscopy, we investigated the mode and the timing of the transformation of the pulmonary capillary bed. During the third postnatal week the parenchymal septa rapidly mature to match adult morphology. Even in adult lungs, however, remnants of the immature status are present: A capillary bilayer is regularly found at the base and the tip of the septa. Our observations support the concept that reduction of intervening tissue, partial fusion of the two capillary networks, and preferential growth lead to the mature vascular arrangement. The fact that true mature interalveolar septa show a denser capillary network than alveolar walls abutting onto pleura, bronchi, or larger vessels is consonant with the fusion theory. Towards the nonparenchyma, the capillary network surrounding every airspace had no counterpart to fuse with. From quantitative data it can be calculated that owing to lung growth, mesh size should increase more than four times between birth and adult age. The adult lung network, however, is denser than the one in young animals. This means that new meshes must be added during growth. We propose that small holes observed in sheet‐like regions of the microvascuiature enlarge to form new capillary meshes. With this mechanism of in‐itself or intussusceptional growth, sprouting of individual capillary segments to increase network size is no longer needed.
Tập 216 Số 2 - Trang 154-164 - 1986