Life Sciences Commons^™

Tooth Tissues Of Certain Sharks And Lungfishes, Yasutomo Iwai-Liao, Yoshikage Higashi, Yoshitaka Tamada

Scanning Microscopy

A comparative odontologic study was conducted on teeth and their supporting tissues from extant sharks, fossil sharks, and living lungfishes using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) X-ray microprobe to investigate evolutionary changes in tooth structure of the fishes.

No histological differences between the extant and fossil teeth of lamnoid sharks were observed. The weight percent of calcium in osteodentin of the fossil shark was higher than that in the extant shark. The weight percents of both P and Ca of tooth tissues in the giant extinct shark showed comparatively low values.

Both the upper and lower …

An Integrated Study Of The Morphological And Gross-Elemental Consequences Of Methyl Mercury Intoxication In Rats, With Particular Attention On The Cerebellum, K. Leyshon, A. J. Morgan

Scanning Microscopy

Methylmercury accumulates in the kidney and liver of rats, but fairly selectively damages the cerebellum, resulting in the clinical symptoms of neurological ataxia after prolonged exposures. Within the cerebellum, morphological examination indicated that the small granule cells beneath the Purkinje layer are especially susceptible to the toxin, showing signs of pyknosis during the phase of locomotory disability, whilst the large Purkinje cells are relatively resistant to cytotoxic injury. Flame photometric and electron probe X-ray microanalysis (EPXMA) of digested samples of the major organs failed to detect any significant changes in the Na, K, Ca, Mg, S and P concentrations of …

Hereditary Hair Changes Revealed By Analysis Of Single Hair Fibres By Scanning Electron Microscopy, B. Forslind, M. K. Andersson, E. Alsterborg

Scanning Microscopy

In many disorders with a genetic background the sparsity of scalp hairs may deter the clinician from trying to extract information from single hair fibres. Presenting a number of diverse conditions, we propose to show that simple measures can be taken in the doctor's office which makes single fibre analysis a useful tool for assessment of factors involved in genetic disorders including the integument and its appendages. The paper is focussed on the utilization of the scanning electron microscope with the goal of demonstrating that pertinent information can be gained where information from transmission electron microscopy and other techniques are …

Elemental Analysis And Fine Structure Of Mitochondrial Granules In Growth Plate Chondrocytes Studied By Electron Energy Loss Spectroscopy And Energy Dispersive X-Ray Microanalysis, Joanna Wroblewski, Romuald Wróblewski, Claudie Mory, Christian Colliex

Scanning Microscopy

Electron energy loss spectrometry - EELS, and energy dispersive X-ray microanalysis - XRMA, were used to study the elemental composition of mitochondrial dense granules - mdg. The study was performed on dry cut thin sections (80-200 nm) of freeze-dried and low temperature embedded cartilage. Results obtained by means of XRMA clearly showed high phosphorus and calcium content in the mdg. Using EELS at 100 kV primary voltage we found that small concentrations of elements (i.e. below typically 1% atomic weight) are difficult to analyze and map, this especially in sections thicker than 50-60 nm. Surprisingly, analysis of calcium can be …

Effect Of A Platinum Chemotherapy Drug On Intracellular Elements During The Cell Cycle, Using X-Ray Microanalysis, P. G. Edwards, M. D. Kendall, I. W. Morris

Scanning Microscopy

Intracellular elemental concentrations were measured in the cytoplasm, nucleus and nucleolus of cultured Chinese hamster ovary (CHO) cells, using energy dispersive electron probe X-ray microanalysis and transmission electron microscopy. Synchronous CHO cell populations were analyzed at different times during the growth cycle. The elements K, P, Mg and Zn were all more concentrated in the nucleus and nucleolus than in the cytoplasm, while no specific subcellular compartmentalization was evident for the elements Na, Cl, Ca, Fe and S. Significant changes in intracellular elemental concentrations were associated with the progression of cells from G₁ phase to S phase of the …

Elemental Mapping Of Cryosections From Cnidarian Nematocytes, Karl Zierold, Pierre Tardent, Sergey V. Buravkov

Scanning Microscopy

The distribution of elements in stinging capsules containing cells called nematocytes is shown by pseudocoloured maps representing the X-ray intensity collected from freeze-dried cryosections. This method provides a distinct overview in addition to the quantitative evaluation of single X-ray spectra. Selected examples illustrate the elemental compartmentation in various cnidarian animals. In particular the matrix of capsules in Hydra vulgaris contains high amounts of K in comparison to the tubule, the surrounding capsule wall and the cytoplasm, whereas in Actinia eguina capsules have either high concentration of Ca or Mg, the latter accompanied by S.

Copper Toxicity In Erwinia Amylovora : An X-Ray Microanalytical Study, N. Hodson, D. C. Sigee

Scanning Microscopy

The effect of Cu²⁺ ions on the growth and elemental composition of the phytopathogenic bacterium Erwinia amylovora was investigated by in vitro culture in nutrient broth supplemented with CuSO₄.

No detectable inhibition in bacterial growth occurred with medium supplemented at 10^-4M Cu (compared to control medium with no added Cu), but partial inhibition occurred at 10^-3M Cu (limiting toxicity) and complete inhibition at 10^-2M Cu.

Under conditions of limiting toxicity, incubation in Cu²⁺ ions leads to a reduction in the general synthesis of cell materials and the formation of abnormally …

Reliability Of Intracellular Water And Ion Distributions As Measured By X-Ray Microanalysis - A Review, Thomas Von Zglinicki

Scanning Microscopy

X-ray microanalysis can be an important tool to reveal the spatial relationships between polyelectrolytes, ions, and water as they occur within cells and tissues in vivo. To reach this goal, at least two of these three closely interrelated variables should be measured independently. Moreover, the absence of systematic errors should be proven.

The present review discusses the probability of artificial ion and water shifts between intracellular compartments due to the growth of dendritic ice crystals much larger than the cross-sectioned remnants commonly seen in frozen-dried sections. Considering the possible mechanism of ice crystal growth it is concluded that ions and …

Cryopreparation Of Tissue For Clinical Applications Of X-Ray Microanalysis, Godfried M. Roomans

Scanning Microscopy

A number of diseases is associated with changes in ion and/or water distribution at the tissue or cell level, and X-ray microanalysis can be used to investigate the relationship between the disease process and the changes in elemental distribution. For analysis of diffusible elements by X-ray microanalysis, the tissue has to be prepared by cryotechniques. To carry out studies of this kind in a clinical environment poses a number of problems. Some of these problems occur already before the tissue is frozen, others are caused by the practical and ethical limitations that are imposed on the freezing method itself when …

Diapause In The Earthworm, Aporrectodea Longa: Morphological And Quantitative X-Ray Microanalysis Of Cryosectioned Chloragogenous Tissue, A. J. Morgan, C. Winters

Scanning Microscopy

The earthworm, Aporrectodea longa, which experiences an apparent obligatory physiological resting state (“diapause”) in the temperate Summer, was collected monthly from February through to October. Morphological examination of the chloragocytes, and quantitative electron probe X-ray microanalysis of their constituent chloragosome granules prepared by cryoultramicrotomy and air-dried/smearing, were undertaken over this period. Worms entered diapause in May, and emerged from it between August and September. The long-term energy stores in the form of lipid, in addition to the more usual polysaccharide (glycogen) reserves, were accumulated by early diapause; the lipid was gradually consumed during diapause. In addition, the structure and composition …

Changes In Sodium Concentration In Cardiac Myocytes From Diabetic Rats, Alice Warley

Scanning Microscopy

The effects of streptozotocin induced diabetes on rats were studied. The animals showed an increase in blood glucose concentration and a loss of weight from both the body and the heart. Loss of weight from the heart was less severe leading to an increased heart to body weight ratio. Study of element concentrations by X-ray microanalysis showed that there was an increase in intracellular Na concentration in cardiac myocytes from the diabetic animals, but no change in Mg. These results agree with studies which show changes in Na/K ATPase after the onset of diabetes.

X-Ray Microanalytical Studies Of Freshwater Biota: Changes In The Elemental Composition Of Anabaena Spiroides During Blooms Of 1988 And 1989, S. Clay, D. C. Sigee, E. Bellinger

Scanning Microscopy

Samples of Anabaena spiroides were collected from a eutrophic freshwater lake (Rostherne Mere, Cheshire) during the bloom periods of 1988 and 1989, and analysed by SEM electron probe X-ray microanalysis. Vegetative cells, akinetes and heterocysts consistently had clear peaks of Mg, Si, P, S, Cl, Kand Ca.

In comparison to vegetative cells, akinetes had generally higher elemental mass fractions (particularly K and P) while heterocysts had significantly lower levels of Mg, P, and Ca.

For both years, elemental mass fractions in vegetative cells showed significant changes during the course of the bloom, but these were not consistent from one year …

The Hall Method In The Quantitative X-Ray Microanalysis Of Biological Specimens: A Review, Godfried M. Roomans

Scanning Microscopy

In the two decades since its inception by T.A. Hall, the continuum theory of quantification has become the general method for quantitative analysis of biological specimens. Although the method was originally developed for thin specimens, its use has been extended to thicker specimens, and it has also been used in quantitative determinations of local water content. The single most important difficulty in the application of the "Hall method" is the accurate calculation of the extraneous continuum, i.e., the continuum due to non-specimen sources. The different variations in methods for quantitative analysis of local water content are critically compared and a …

Cryofixation Of Heart Tissue For X-Ray Microanalysis, Alice Warley

Scanning Microscopy

Cryofixation of tissues is necessary to be able to study the concentrations of elements by X-ray microanalysis. Simple dissection of heart tissue fragments of the size needed for optimum cryofixation by the conventional methods of plunge or slam freezing leads to the development of ischaemia in the tissue fragments and a consequent redistribution of the diffusible elements. Heart tissue can be frozen in vivo using liquid nitrogen cooled Cu clad pliers, but the morphological detail is preserved better if the cooled pliers are exposed to liquefied propane immediately before freezing, Concentrations of Na are lower and concentrations of K are …

Strontium And Bromide As Tracers In X-Ray Microanalysis Of Biological Tissue, Joanna Wroblewski, Samuel Sagström, Helmy Mulders, Godfried M. Roomans

Scanning Microscopy

Since energy dispersive X-ray microanalysis cannot distinguish between isotopes of the same element, alternative methods have to be used to get information similar to that obtained in experiments with radioactive tracers. In the present study, strontium was used as a tracer for calcium, and bromide as a tracer for chloride. Rats were injected with strontium chloride in vivo, and the uptake of strontium in the acinar cells of the submandibular gland was studied. Eventually a Sr/Ca ratio of 0.3 was reached. In some animals, secretion of mucus had been elicited by stimulation with isoproterenol 4 h prior to injection …

Microscopy Analysis Of Dental Titanium Casting Investment Materials, H. J. Mueller

Scanning Microscopy

Scanning electron microscopy (SEM) analysis and qualitative energy dispersive x-ray microanalysis (EDS) of investment materials for dental titanium casting were performed. Two commercial investment materials, Ohara and Castmatic, and an experimental zirconia material were analyzed. The microstructural constituents and the unfired and fired structures were included. Larger refractory particles and matrix embedding smaller particles were observed with each material. Detection of aluminum, silicon, magnesium, zirconium and oxygen provided a basis to reason the presence of alumina (Al₂O₃), silica (SiO₂), magnesia (MgO), and zirconia (ZrO₂). Hence, Ohara contained quartz and an alumino-silicate, Castmatic …

Biuret Staining And X-Ray Microanalysis For Locating Grafted Poly(Methyl Acrylate) On Wool Fibers, R. R. Bresee, P. A. Annis

Scanning Microscopy

Biuret reagent was used to stain wool fibers with copper so the location of a small amount of poly(methyl acrylate) grafted onto the fibers could be determined by energy dispersive x-ray microanalysis of copper. The grafted polymer was determined to be located in regions of the fibers where cuticle had been previously damaged. The amount of grafted polymer present was too small for secondary electron imaging to be useful for locating the polymer grafts.

X-Ray Microanalysis Of Calcium Containing Organelles In Resin Embedded Tissue, G. Nicaise, I. Gillot, A. K. Julliard, E. Keicher, S. Blaineau, J. Amsellem, J. C. Meyran, M. L. Hernandez-Nicaise, B. Ciapa, C. Gleyzal

Scanning Microscopy

The localization of calcium in cell organelles at the electron microscope level is often achieved through cytochemical techniques, and verified by X-ray microanalysis. Various methods have been used to cytochemically detect calcium or calcium-binding sites : calcium loading, calcium substitution by strontium, barium, or even lead, and calcium precipitation by oxalate, phosphate, fluoride, or pyroantimonate. Their results may have heuristic value, particularly in preliminary studies of poorly known cell types. A complementary and more physiological approach is offered by quantitative measurement of the total calcium content of organelles after cryofixation.

Resin embedding is less demanding than cryomicrotomy and gives better …

Location And Identification Of Colloidal Gold Particles On The Cell Surface With A Scanning Electron Microscope And Energy Dispersive Analyzer, Sinikka Eskelinen, Raija Peura

Scanning Microscopy

The use of colloidal gold particles for locating cell surface components by scanning electron microscopy (SEM) has been restricted due to difficulties in the identification of these gold particles under SEM. It is shown here how the gold particles bound to cell surfaces can be located and identified under SEM using the secondary electron imaging (SEI) mode with an energy dispersive X-ray microanalyzer (EDS). This enables reliable identification of gold particles and good quality micrographs of the cells to be achieved at the same time. The distribution of receptors for two lectins, concanaval in A (ConA) and wheat germ agglutinin …

Preparation Of Cultured And Isolated Cells For X-Ray Microanalysis, K. Zierold, D. Schäfer

Scanning Microscopy

Various electron microscopical preparation techniques are compared with regard to the preservation of the intracellular element distribution as determined by X-ray microanalysis in scanning and scanning transmission electron microscopy. By use of chemical agents for fixation and dehydration ions are redistributed and washed out. This is also true for freeze-substitution. Whole cells are prepared by cryofixation followed by freeze-drying. Interference of intracellular measurements by extracellular elements can be avoided by appropriate washing the cells before cryofixation. The washing medium has to be carefully selected in order to avoid distortions of the original intracellular element content. These problems are circumvented by …

Intracellular Water Distribution And Aging As Examined By X-Ray Microanalysis, Thomas Von Zglinicki

Scanning Microscopy

The results reviewed here demonstrate that 1. the distribution of dry mass as observed in frozen-dried cryosections might be used as an unbiased measure of intracellular dry mass resp. water distributions in the tissue in vivo and 2. the well-known loss of water from cells during aging is solely due to a water loss from mitochondria without changes in the water content of all other components of the cell in the case of rat liver and heart muscle.

The reason for the water loss might be increased counter ion binding by membrane-bound enzymes due to decreased fluidity of the inner …

Extraneous Background-Correction Program For Matrix Bound Multiple Point X-Ray Microanalysis, W. C. De Bruijn, M. P. C. Van Miert

Scanning Microscopy

A program is described that allows online determination of extraneous background in multiple point X-ray microanalytical matrices. The program is based upon the calculations of the extraneous background for the film (when present), the standard and the unknown by (100 sec.) point analysis. The program searches for a peak-free part of the spectrum in which the calculated value for the extraneous background is about equal to the value in this region of the spectrum (=b_e). Online the contents of this b_e-region is subtracted from an unmanipulated continuum region in the vicinity of the element present in …

Elemental Levels In Mast Cell Granules Differ In Sections From Normal And Diabetic Rats: An X-Ray Microanalysis Study, Marion D. Kendall

Scanning Microscopy

Mast cells around the thymus of rats stain red with alcian blue and safranin indicating that the mast cells are probably of the peritoneal (connective tissue) type. After the onset of streptozotocin induced diabetes some cells contain both red and blue granules and blue staining cells may appear.

X-ray microanalysis of frozen freeze-dried sections from diabetic male CSE Wistar rats showed electron dense granules to have similar amounts of S to normal rat mast cell granules but reduced levels of Na, Mg, P, Cl and K. Two cells also had electron lucent granules with very high levels of Na, Cl, …

Scanning Electron Microscopy, X-Ray Microanalysis And Immunohistochemistry On Worn Soft Contact Lenses, Piera Versura, M. C. Maltarello, Godfried M. Roomans, R. Caramazza, R. Laschi

Scanning Microscopy

The deposits accumulated on the surfaces of soft contact lenses are a cause of problems for the wearer of these lenses, as the deposits are never completely removed by the available washing solutions. Therefore it appears of interest to investigate the composition of these deposits.

In this paper we review the major findings in the literature and, in addition, present our personal experience.

We have studied new, continuously and daily worn soft contact lenses by scanning electron microscopy (SEM), X-ray microanalysis and immunohistochemistry. We have carefully evaluated preparative methods, and we can conclude that SEM and X-ray microanalysis are best …

Image Analysis And X-Ray Microanalysis In Cytochemistry, W. C. De Bruijn, H. K. Koerten, M. I. Cleton-Soeteman, C. J. G. Blok-Van Hoek

Scanning Microscopy

When cytochemical reaction products are homogeneously distributed within an organelle, point analyses suffice for the quantitative approach. However, quantitative analysis becomes tedious, when the elements in the reaction product are inhomogeneously distributed. Problems arise when elements from two reaction products have to be related to each other, or to endogenous cytological products (ferritin, haemosiderin, calcium, electron dense markers), either topographically or in concentration. When analyzing inhomogeneous/heteromorphical reaction product-containing organelles special attention has to be paid to measure and relate both volume and concentration. In this paper a relative simple structure (eosinophil granules) is chosen to demonstrate that the acquisition of …

Fictitious Calculi And Human Calculi With Foreign Nuclei, P.-T. Cheng, K. P. H. Pritzker, J. Richards, D. Holmyard

Scanning Microscopy

The correlative approach employing polarized light microscopy, x-ray powder diffraction, scanning electron microscopy, transmission electron microscopy, selected area electron diffraction and energy dispersive x-ray microanalysis proves to be very useful in identifying fictitious calculi and genuine human calculi with foreign body nuclei. The common artifacts as reported in the literature and observed also by us were minerals, vegetable and plant seeds, cereals, sand grains and sea shell fragments.

Two interesting cases involving foreign body nuclei have been reported: one urinary calculus containing a piece of plastic-coated titanium foil in the center; one nasal calculus with a nut as a nucleus. …

X-Ray Microanalysis Of Cells In Suspension And The Application Of This Technique To The Study Of The Thymus Gland, A. Warley

Scanning Microscopy

Although X-ray microanalysis has become a well established technique in the study of tissues there are still relatively few papers reporting results from cells in culture. Our early investigations on freshly isolated murine thymocytes reported higher than expected concentrations of Na and Cl in these cells but subsequent studies have shown that these high concentrations are artefactual. Rat thymocytes that have been isolated and incubated for a short time to allow them to recover from the isolation procedure have lower concentrations of Na and Cl. Sections prepared from pellets of cells which have been concentrated by centrifugation in a microhaematocrit …

Studies On The Occurrence And Elemental Composition Of Bacteria In Freshwater Plankton, K. N. Booth, D. C. Sigee, E. Bellinger

Scanning Microscopy

The occurrence and cation content of bacteria in a eutrophic freshwater lake (Rostherne Mere, Cheshire, UK) were investigated over a one year sampling period in relation to cation changes in the lake surface water and phytoplankton.

Scanning electron microscope examination of trawl-net and filtered samples demonstrated bacterial association with Anabaena, Aphanizomenon and diatoms. Direct counts of associated and unassociated bacteria showed that increases in bacterial population relate to population decline of major algal constituents.

Spectrophotometric determination of selected cation levels in the lake water demonstrated wide fluctuations throughout the sampling period, with elevated levels of transition metals before and at …