Morphometric Analysis Techniques Research Articles

Assessment of endometrial differentiation and function

The failure to produce an endometrium which is receptive and able to support implantation may underlie some cases of infertility and represent a contributory factor preventing higher pregnancy rates in IVF. This is of direct relevance to ovarian hyperstimulation protocols which may affect the pattern of endometrial differentiation and function and to patients possessing intrinsic defects in their endometrial response. It is apparent that markers of endometrial differentiation are required, particularly those defining the ability of the tissue to support implantation. The common method of assessing endometrial differentiation by histological evaluation, which has been substantially improved by techniques of morphometric analysis, depend upon alterations in endometrial structure. However, the production of monoclonal antibodies to proteins and epitope-bearing mucins which exhibit specific patterns of expression during the luteal phase now also enables immunohistological assessment of endometrial differentiation. Whether these proteins mediate essential functions is not known but as they represent end-points of hormone action this would suggest that their measurement may form the basis of tests of endometrial function and supplement morphometric analyses.

A morphometric and x-ray energy dispersive approach to monitoring pH-altered cadmium toxicity inAnabaena flos-aquae

Cadmium toxicity and uptake as influenced by different pH values have been studied in the freshwater cyanobacterium Anabaena flos-aquae, using the techniques of morphometric analysis, x-ray energy dispersive analysis and atomic absorption spectrophotometry. A general reduction in cell dimension, thylakoid surface area, number and volume of polyhedral bodies, polyphosphate bodies, cyanophycin granules, lipid bodies, membrane limited crystalline inclusions, volume and number of wall layers and mesosomes was observed. These reductions were more pronounced in both acidic and alkaline medium than at pH 7.2. At 0.12 microM Cd, the uptake increased with alkaline pH values, and uptake was greater at pH 7.2 than at either acid or alkaline pHs. Lysis of cell wall at 1.18 microM Cd showed the following decreasing trend: pH 4.0 greater than pH 5.5 greater than pH 10.0 greater than pH 9.0 greater than pH 7.2. There was a total loss of lipid bodies at 1.18 microM Cd at all pH values listed. It is suggested that these techniques can be successfully employed for bioassay studies of metal toxicity to algae. In particular, cell wall lysis and loss of lipids by algae are good indicators of pH effects and metal toxicity in the aquatic ecosystem.

CRUSTACEAN BRAIN LIPOFUSCIN: AN EXAMINATION OF THE MORPHOLOGICAL PIGMENT IN THE FRESH-WATER CRAYFISH CHERAX CUSPIDATUS (PARASTACIDAE)

ABSTRACT Morphological lipofuscin was specifically demonstrated in the central nervous system of a crustacean for the first time. It was primarily associated with neuroglia and axons within cell masses in the brain, particularly the olfactory nuclei. It exhibited fluorescence, staining, and ultrastructural properties similar to the lipofuscin of other invertebrates and vertebrates. Pigment was quantified using morphometric image analysis techniques and shown to be more concentrated in larger animals than smaller ones. The conclusive demonstration of the presence of lipofuscin in a crustacean brain should facilitate the refocusing of crustacean age-determination studies.

Comparative Morphometric Analysis of Cochlear Vessels in Wistar‐Kyoto Rats, Spontaneously Hypertensive Rats, and Aged Spontaneously Hypertensive Rats

Vessel density and the ratio of the tissue area to the vessel surface area were studied by morphometric analysis techniques in normal rats, spontaneously hypertensive rats (SHR), and aged spontaneously hypertensive rats (aged SHR). Horseradish peroxidase was injected intravenously and the animals were killed 10 minutes later. The temporal bones were harvested, fixed in glutaraldehyde, and decalcified in 10% ethylenediamine tetraacetic acid (EDTA). After 7 days of decalcification, the cochleas were dissected and incubated with a diaminobenzidine tetrahydrochloride solution. Sections with stained vessels were projected onto the digitizing plate with the help of the camera lucida. The computer was used to calculate tissue area, vessel length, and vessel surface area. A statistically significant increase (p less than 0.05) in both the tissue area to vessel length ratio and the tissue area to vessel surface area ratio was demonstrated in the SHR and the aged SHR groups when compared to the WKY in the stria vascularis. No statistically significant difference was found between the two SHR groups. These data show a decrease of the vessel density in the capillary beds of the stria vascularis in spontaneously hypertensive rats. No statistically significant difference was found in the diameters of the capillary among the three groups.

A Morphometric Study on the Effects of Ethylene Treatment in Promoting Abscission of Tobacco Flower Pedicels

The ultrastructural changes observed in ethylene-induced abscission of tobacco flower pedicels (Nicotiana tabacum L. ;Little Turkish') were studied by the techniques of morphometric analysis. The surface area of the membranes, relative volume of the organelles, and the number of organelles were determined for both ethylene-treated and control cells. In pedicels exposed to ethylene for 4.5 to 5 hours, abscission was evident within the separation zone. The most significant change in cell structure was observed in the surface area of the rough endoplasmic reticulum which more than doubled with ethylene treatment of the tissue.

Utilization of morphometric analysis in evaluating response of Plectonema boryanum (Cyanophyceae) to exposure to eight heavy metals.

The techniques of morphometric analysis were used to study the ultrastructural changes induced inPlectonema boryanum (Cyanophyceae) by heavy metal exposure. Four of the test metals (Mn, Zn, Hg, and Cd) produced no changes in overall cell dimensions. Two metals (Pb and Cu) produced significant increases in cell size and two other metals (Co and Ni) caused significant decrease in cell size. Mn, Pb, Co, Ni, and Cd caused significant increases in the surface area of the cell's thylakoids and Ni, Co, Zn, and Hg caused a significant reduction in the volume of the intrathylakoidal spaces. All tested metals except Pb and Mn caused the production of extra intracellular membrane whorls. Pb, Mn, Co, Zn, Hg, Cd, and Ni caused coalescence of cellular lipid while Cu caused a reduction in cellular lipid content. Co caused a reduction in the number of polyhedral bodies and Cd caused an increase in the volume of the cell's polyphosphate bodies. The physiological significance of these findings are discussed in terms of potential toxic action of cations and the cellular mechanisms for the detoxification of cations once they enter the cell. Common mechanisms of both procaryotic and eucaryotic algae are discussed and the utilization of stereological techniques for electron microscopic morphometric analysis is established in studies of metal toxicity at the cellular level.

Fluorophosphate-sensitive esterases in mammalian liver: the radioautographic localization and measurement of fluorophosphate-reactive sites in adult rat liver.

Fluorophosphate-reactive (FPR) sites in the adult male rat liver, tentatively identified as esterase active centers, were localized and measured using the combined techniques of quantitative electron microscope radioautography and morphometric analysis with the light and electron microscope. The FPR sites were measured in liver which had been prefixed by perfusion with 1.5% glutaraldehyde and reacted with 10(-4) M tritium-labeled diisopropyl fluorophosphate (3H-DFP). Under the experimental conditions 64-67% of the esterase activity in fresh liver was retained for reaction with the 3H-DFP, which is known to bind irreversibly to the active sites of certain esterases. In light and electron microscope radioautographs the developed silver grains were concentrated over the cytoplasm of hepatocytes. A low concentration occurred over erythrocytes. All other areas in the liver had a concentration of grains resembling the background concentration. Quantitative measurements of grain density in the electron microscope radioautographs revealed the highest density, after correcting for radiation spread, in cytoplasmic granules (mainly cytolysomes). The grain densities over the rough and smooth endoplasmic reticulum and associated structures were also equal to or above the average hepatocyte grain density. Due to the large fractional volume of endoplasmic reticulum per hepatocyte (58% of cell volume) and the fraction of the liver occupied by hepatocytes (79% of liver volume) the majority of FPR sites in the liver occurred in the rough and smooth endoplasmic reticulum and associated structures. The average numbers of FPR sites were calculated per micrometer3 of hepatocyte (5.0 x 10(5) sites/micrometer3) and per unit volume of each significantly labeled organelle. In addition, the number of FPR sites per hepatocyte (2.5 X 10(9) sites/cell), per cm3 liver (4.1 X 10(17) sites/cm3) and in the total liver of an average 100 g male rate (2.2 X 10(18) sites/total liver) were also calculated.