The influence of upstream predation on the expression of fish effects in downstream patches

1. Enclosures were installed in a fishless stream and divided transversely into upstream and downstream sections. Downstream sections were further divided longitudinally, and one of the downstream sections in each enclosure was stocked with juvenile coho salmon ( Oncorhynchus kisutch ), and the other side was left as a fishless control. Densities of juvenile coho were then manipulated in upper sections of enclosures to determine the effect of upstream predation intensity on fish predation effects in downstream sections.
2. Significantly fewer mayflies (primarily Ameletus sp.) were observed grazing on unglazed ceramic tiles in lower enclosure sections with fish present. There was no detectable effect of fish density in upstream enclosure sections on the number of mayflies observed grazing on ceramic tiles in lower sections of the enclosures.
3. There was a significant positive effect of both fish presence and upstream density on chlorophyll a concentrations on ceramic tiles in lower enclosure sections, but not on chlorophyll a on natural gravel substratum.
4. Behavioural experiments with mayflies and coho in streamside troughs suggest that Ameletus sp. responds primarily to mechanical rather than chemical cues from coho parr.     

Split Nucleoli as A Source of Error in Nerve Cell Counts

The number of nerve cells in a given ganglion or nucleus is usually determined by counting the nucleoli in serial sections. The possibility that nucleoli may split and appear in more than one section is recognized as a source of error. A determination of the value of this error was made as follows; from nodose ganglia of four cats were cut serial transverse sections in which the sections varied in thickness. Thus from one ganglion, four sections were cut at 12 μ, then six at 9μ, and eight at 6μ. The process was repeated over and over until the entire ganglion was sectioned. The other ganglia were sectioned similarly. After mounting and staining, separate counts were made of the nucleoli of each given ganglion from the sections of different thicknesses. If nucleoli split according to theoretical expectations, the percentage of nucleoli split in thick sections should be less than the percentage split in thinner sections and the counts based on the sections of different thicknesses should vary accordingly. The results obtained indicate that the counts from thin sections do not differ appreciably from counts from much thicker sections, i. e., the thickness of the sections does not affect the count. It is, therefore, concluded that no correction should be made for split nucleoli if the sections are around 10 μ in thickness and none but distinct and definite nucleoli are counted.     

Characteristics of serial cross-sections of hairs of the Japanese monkey (Macaca fuscata fuscata)

The characteristics of serial cross-sections of hairs collected from an adult male Japanese monkey were investigated. Cross-sections were made of five to eight pieces per hair. The shapes of the cross-sections were elliptical or rounded on the whole. The fibre indices of the sections ranged from 83 to 100. In particular, those of proximal (basal) sections were close to 100. The hair diameter was 86.4 μ at maximum and 27.2 μ at minimum. A tendency was observed for the longer hairs to have thicker diameters. The changes in thickness along the fibre shaft were slightly different in relation to hair length. The thickest point was at around the middle of the fibre in the intermediate hair, somewhat towards the top of the central part in the long hair, and somewhat towards the base in the short hair. The hair of the Japanese monkey, however, was considered to be scanty in changes along the fibre shaft in comparison with many other animals. Medullae could scarcely be seen in the short hair and in the terminal and proximal sections of all hairs. Their shapes in cross-section were not uniform and rough at the margins. The fibre-medulla indices were generally less than 30 and smaller than those of many other mammals. Pigmentary granules were observed in all sections examined. The granules were black-grey in sections of the black-grey coloured part and yellow in the yellowish sections. They were dense in distal sections and scarce in sections close to the base. The cross-sectional appearance of the thickest part of the long hair was considered to be useful for hair identification, since it was good in pigmentation and medullation and relatively small fibre index.     

激光微切割与定量PCR技术分析肾脏病理切片RNA

采用激光微切割与定量PCR技术,分析使用不同提取方法从不同固定方法固定的病理切片中提取的RNA.用70%乙醇、丙酮、甲醇、4%多聚甲醛固定肾脏冰冻切片,使用激光微切割技术切取肾小球,用硫氰酸胍方法(guanidinethiocyanatemethods,GTC)和Trizol试剂方法提取RNA,使用Taqman定量PCR方法分析比较各组RNA的量;选取丙酮固定的石蜡切片,使用激光微切割技术切取肾小球,采用RNA裂解液提取RNA,使用Taqman定量PCR方法,比较石蜡切片和冰冻切片中RNA含量.结果显示:提取沉淀性固定剂如乙醇、丙酮、甲醇固定的冰冻切片的RNA时,2种提取方法和3种固定方法对RNA含量的影响都无明显差异;但在提取4%多聚甲醛固定冰冻切片时,使用Trizol提取RNA含量明显高于使用GTC方法,且其含量与沉淀性固定剂固定的切片RNA含量无明显差异.石蜡切片中经激光微切割肾小球的RNA含量与冰冻切片经激光微切割肾小球的RNA含量无明显差异.结果提示:切片的固定方法和RNA的提取方法是影响切片RNA提取量的主要原因.     

A comparative study of morphometric measurements of nucleoli in uveal melanomas from electron micrographs and plastic-embedded and paraffin-embedded sections

Morphometric measurements of nucleoli were done on uveal melanomas from surviving and nonsurviving patients. The melanomas were embedded in paraffin and plastic, and measurement data from Papanicolaou-stained paraffin-embedded sections, toluidine blue-stained plastic-embedded sections and scanning transmission electron micrographs (STEM) of plastic-embedded sections were compared. The results showed that one parameter, the coefficient of variation (CV) of nucleolar area, correctly classified 80% of the cases as to survival when plastic-embedded material was used and 70% of the cases when paraffin-embedded material or STEM micrographs were used. The inverse standard deviation of the nucleolar area was a better predictor of outcome than was the CV of nucleolar area only in the paraffin-embedded sections. The nucleolar measurements were most easily and rapidly performed in the plastic-embedded sections.     

声带组织切片方法的比较

目的:探讨犬声带冠状位切片与水平位切片各自的特点,为声带实验提供合适的切片方法。方法:家犬4只,2只取材后行冠状位石蜡切片,2只取材后行水平位石蜡切片。通过HE染色观察声带固有层的一般组织结构,Masson三色染色观察固有层中胶原的排列情况。结果:HE染色示冠状位、水平位切片均可见声带表面被覆复层鳞状上皮,固有层内有大量排列紧密的纤维组织,纤维组织中夹杂少量腺体,固有层下方为肌层。冠状位切片可观察声带某一点冠状面固有层的情况,若观察整个声带的情况需声带连续切片;水平位切片可在一张切片中观察到前联合、声带膜部及声带突部位的固有层情况,解剖标志明显,利于定位。Masson三色染色示冠状位、水平位切片均可见固有层浅层有较细的胶原纤维束,中层有较粗的纤维束与较细的纤维束交织排列,深层纤维束排列更紧密。结论:冠状位切片可观察声带某一点冠状面固有层的整体情况,水平位切片可在一张切片中观察到前联合、膜部及声带突部位的固有层情况。     

Protein Composition in Relation to Age of Daffodil Leaves

Daffodil foliage leaves were divided into sections along theirlength; the basal sections then contained the youngest, growingregions of the leaves, and the other sections represented progressivelyolder tissue as the leaf apex was approached. Representativeprotein fractions were isolated from some of these sections,and after hydrolysis their amino-acid compositions were compared.Protein from bulb scale leaves was also analysed. Within thefoliage leaf, age did not markedly affect the composition ofthe proteins. Larger differences of composition were found whenthe proteins of the bulb scale, a typical storage tissue, werecompared with those of the foliage leaves. The free amino-acid complements of the different sections ofthe foliage leaves were also compared. Variation of compositionwith leaf age did occur, but no generalizations can be madethat are applicable to all amino-acids.     

Staining Sulfated Mucopolysaccharides in Sections by Means of Acriflavine

Acriflavine gave insoluble salts with sulfated esters. Frozen or paraffin sections (fixed in 10% formol or Carnoy's solution) were stained in M/20 acriflavine solution and excess dye was rinsed in 95% alcohol. Then nuclei were stained with Meyer's haemalum. Thereafter the sections were washed in water, dehydrated in alcohol, cleared in xylene and mounted in balsam. Sulfated esters in the tissue sections were colored yellow or orange-yellow, generally more densely in frozen than in paraffin sections.     

Application of glial fibrillary acidic protein immunohistochemistry in the quantification of astrocytes in the rat brain

Immunohistochemical staining for glial fibrillary acidic protein (GFAP) was employed as a tool for quantification of astrocytes in the rat brain. One-micron-methacrylate sections were prepared from 70-micron slices stained for GFAP by using a preembedding staining procedure. Numbers/unit area of astrocytes and nonastrocytes were determined for cortex, corpus callosum, and hippocampal neuropil. In each, counts from GFAP-stained, toluidine-blue-counterstained sections were compared with counts obtained from sections stained with toluidine blue alone. Numbers of nonastrocytes and total glia in all three regions were comparable in both groups of sections. Astrocyte counts in the cortex and hippocampus also showed no significant differences between the two groups. In contrast, the number of astrocytes in the corpus callosum was significantly lower in GFAP-stained, toluidine-blue-counterstained sections than in sections stained with toluidine blue alone. GFAP immunohistochemistry is a useful tool for the quantification of astrocytes in semi-thin plastic sections of rat brain.     

Effects of time and temperature during attachment of sections to microscope slides on immunohistochemical detection of antigens.

To study the effects of time and temperature on attachment of tissue sections to microscope slides, we examined the intensity of immunohistochemical staining of selected antigens in nine different neoplastic and normal tissues after attaching sections at different times and temperatures. Typically, both the temperature and time are minimized when tissue sections attached to slides; however, suboptimal times and temperatures during attachment may result in either loss of tissue due to poor attachment or the necessity for inconvenient staining regimens. Using standard immunohistochemical techniques, 5 microm tissue sections were attached at 58 degrees C for 1, 4 and 24 hr. In a separate study, 5 microm tissue sections were attached for 16 hr at 58, 68 and 80 degrees C. The intensity of staining decreased slightly when the tissue sections were heated at 80 degrees C for 16 hr, but there was little or no decrease when tissues were heated at 68 degrees C or lower for 16 hr, or at 58 degrees C for up to 24 hr.     

Fractionation of proteins of pea stem sections during root formation and its inhibition by kinetin and ethionine

Changes in the protein constituents in pea stem sections during root formation and its inhibition by kinetin and ethionine were studied. Only quantitative differences in the protein fractions separated on DEAE-cellulose column were noted. The formation of foci of meristematic cells in pericycle was accompanied by an increase in the amount of fraction “l”. This fraction disappeared rapidly in sections where root formation either did not occur (internodial sections without nodes) or was inhibited by ethionine (stem sections with basal and apical nodes). Incubation of stem sections in a kinetin solution for 16 hours after cutting of stems partially preserved fraction “l”. The increase in the amount of fraction “l” was one of the first metabolic changes in root-forming pea stem sections after cutting of stems.     

Quantitative histology of the rat thyroid. Influence of histologic techniques on the morphometric data

The influence of various histologic techniques on the results obtained by morphometric analysis of the rat thyroid gland was studied. The limits of thyroid follicles were more clearly defined in both silver-impregnated paraffin-embedded sections and resin-embedded semithin sections than in routinely stained paraffin-embedded sections, thus enabling more accurate measurements of thyroid structures. Due to its simplicity, the silver impregnation method is clearly useful for histomorphometric studies when large numbers of measurements are involved. C cells were easily identified in paraffin-embedded sections by immunohistochemical staining. The measurement of interstitial tissue in sections without immunostaining of C cells led to an overestimation of the volume fraction of interstitial tissue.     

Distribution of acid phosphatase, beta-glucuronidase, n-acetyl-beta-d-glucosaminidase and beta-galactosidase in cornea of albino rabbit.

Activities of acid phosphatase, beta-glucuronidase, N-acethyl-beta-D-glucosaminidase and acid beta-galactosidase were investigated histochemically in rabbit corneas. Frozen sections after block fixation in cold 4% formaldehyde with 1% CaCl2 followed by washing in cold physiological saline as well as cold microtome sections of corneas quenched in petroleter chilled with acetone-dry ice mixture, transferred to nonprecooled slides or semipermeable membranes were used. Standard aqueous media were employed in the case of free-floating frozen sections of fixed corneas as well as of cold mictrotome sections (postfixed in cold 4% formaldehyde). Agar media were used in connection with the technic of semipermeable membranes. Gomori method (in the case of acid phosphatase), simultaneous azocoupling methods (substrates derivated of naphthol-AS-BI with hexazonium-p-rosanilin) in the case of acid phosphatase, beta-glucuronidase and N-acetyl-beta-D-glucosaminidase and the indigogenic method in the case of acid beta-galactosidase were applied. Enzyme activities in sections of fixed corneas were minimal in comparison with those in cold microtome sections of unfixed material revealed particularly with the technic of semipermeable membranes which is to be preferred. This technic is recommended in studies concerned with lysosomal enzymes in the cornea, particularly in keratocytes. All enzymes investigated were present in corneal epithelium, keratocytes and endothelium. Acid phosphatase displayed the highest activity followed by beta-glucuronidase and acetyl-beta-D-glucosaminidase. The activity of beta-galactosidase was the lowest. For the demonstration of activities in keratocytes sections parallel to the surface are very suitable. In these sections enzyme activities were demonstrated in small granules (apparently lysosomes) present in the central part of their cytoplasm as well as in projections. Diffuse staining was also seen, being the highest in the case of acid phosphatase.     

A New Method for Preparation of Undecalcified Bone Sections

A new method for preparation of sections of undecalcified bone is described. Samples of ovine bone were embedded in methylmethacrylate and thick-sectioned with a cutoff machine or commercial band saw. Composite slides were prepared by gluing white acrylic to glass using cyanoacrylate glue. Bone sections were glued to the composite slide and then surface polished by grinding or ultramilling. The polished surface of the section was then etched and stained. The techniques described in this paper reduce the time spent grinding or milling sections and improve resolution of surface-stained features of undecalcified bone sections.     

Biosynthesis of avenacins and phytosterols in roots of Avena sativa cv. Image

In keeping with the proposal that avenacin biosynthesis is restricted to the tips of primary roots of oat seedlings, the incorporation of radioactivity from R-[2-(14)C]mevalonic acid (MVA) into avenacins and beta-amyrin by serial sections of primary roots was found to be more-or-less restricted to root tip sections. Squalene synthase (SQS) (EC 2.5.1.21) and 2,3-oxidosqualene:beta-amyrin cyclase (OS beta AC) (EC 5.4.99) were also most active in these sections. The incorporation of radiolabel from R-[2-(14)C]MVA into cycloartenol and 24-methylene cycloartanol by, and the 2,3-oxidosqualene:cycloartenol cyclase (OSCC) (EC 5.4.99) activity in, the various serial sections were consistent with phytosterol biosynthesis occurring in all the sections of the root with some tailing-off in the rate of synthesis in the more distal sections.     

The Preparation of Sections of Mineralized Tissue Suitable for the Demonstration of Alkaline Phosphatase

Blocks of molar teeth and bisected knee joints from rats of 7-21 days were embedded, without previous decalcification, in tropical grade ester wax. Serial sections were cut at settings of 3-25μ on a base sledge microtome equipped with a Jung extra hard steel knife with a tool-edged profile. The sections were supported with Sellotape during actual cutting and were then coated with a 2% celloidin solution. Chloroform was used to free the sections from the Sellotape. The distribution of alkaline phosphatase activity in the knee joints and teeth was demonstrated in these sections with the coupled-azo dye technique of Gomori, using Brentamine fast red T.R. salt.     

In vitro adherence of lymphocytes to unfixed and fixed high endothelial cells of lymph nodes.

Rat thoracic duct lymphocytes (TDL) bind selectively to venules lined by high endothelial cells (HEV) when overlaid onto glutaraldehyde-fixed frozen sections of lymph nodes. This report describes the characteristics of TDL binding to HEV in unfixed frozen sections and compares this reactivity with that observed after fixing sections with different reagents. We found that TDL bound to unfixed HEV and that the pattern of adherence to such sections was identical to that observed when using glutaraldehyde-fixed tissue. Fixation of the sections with glutaraldehyde, however, enhanced the binding reaction. This effect was also observed when sections were treated with the diimidoester, dimethylsuberimidate (DMS) but not when methanol or formaldehyde was used. Since glutaraldehyde and DMS are each bifunctional cross-linking reagents, the results suggest that in vitro HEV adherence was facilitated under conditions in which the endothelial binding sites were present in an aggregated form.     

A new method for preparation of undecalcified bone sections

A new method for preparation of sections of undecalcified bone is described. Samples of ovine bone were embedded in methylmethacrylate and thick-sectioned with a cutoff machine or commercial band saw. Composite slides were prepared by gluing white acrylic to glass using cyanoacrylate glue. Bone sections were glued to the composite slide and then surface polished by grinding or ultramilling. The polished surface of the section was then etched and stained. The techniques described in this paper reduce the time spent grinding or milling sections and improve resolution of surface-stained features of undecalcified bone sections.     

The effect of ancymidol on hyperhydricity, regeneration, starch and antioxidant enzymatic activities in liquid-cultured Narcissus

 Addition of the growth retardant ancymidol to Narcissus shoots and lower inner leaf sections isolated from shoots cultured in liquid medium induced hyperhydric malformations associated with morphogenetic changes. Meristematic centers initiated on the basal proximal ends appeared over the entire surface of the hyperhydric leaf sections after 6 weeks in culture. The meristematic centers which formed clusters on the leaf sections developed later into buds. In leaf sections grown in the liquid medium lacking ancymidol, hyperhydricity was not induced, and regeneration was not observed. Starch and protein levels and ascorbate peroxidase and catalase activities were examined in shoots and isolated leaf sections that were either hyperhydric or non-hyperhydric. In ancymidol-treated, hyperhydric leaf sections, ascorbate peroxidase and catalase activities were lower than in control, untreated leaf sections. The changes in starch and protein levels and in antioxidant enzymatic activities appeared to be related to the onset of meristematic-center initiation and further bud development on Narcissus hyperhydric leaf sections. Received: 6 May 2000 / Revision received: 21 August 2000 / Accepted: 22 August 2000     

Improved correction of quantitative nuclear DNA (ploidy) measurements in tissue sections

OBJECTIVE: To evaluate, using a computer model, the advantages for ploidy measurements of selecting only center-containing sections of nuclei in ultrathin, very thick or relatively thick tissue sections. STUDY DESIGN: The computed corpuscle sectioning program was run on a personal computer. Its synthetic data were corrected by a variety of correction algorithms. RESULTS: When only center-containing sections of nuclei were selected in ultrathin sections, spherical nuclei could be corrected perfectly, and mildly prolate ellipsoidal nuclei with a selection bias in favor of elliptical nuclear section profiles could be corrected with high fidelity. Ultrathin sections most faithfully represented the true height of the peak of highest ploidy and showed better peak discrimination than other choices of section thickness, but small sample size, wavy sections, markedly inhomogeneous intranuclear DNA distribution and oblate ellipsoidal nuclei represented significant limitations of this approach. As nuclear prolation increased, peak definition worsened, and the peak of highest ploidy was falsely shortened. Results were unaffected by errors in the estimation of section thickness when an internal diploid standard was used. The effect of variable internuclear DNA concentration in mildly or moderately prolate ellipsoidal nuclei was nil. The choice of correction algorithm was unimportant, except that the reference curve method was better able to analyze oblate ellipsoidal nuclei, wavy sections and nuclei with inhomogeneous intranuclear DNA, and provided superior insight into nuclear and section parameters. Thick and very thick sections did not require correction and, unlike ultrathin sections, were immune to markedly inhomogeneous intranuclear DNA distribution, to nonspherical nuclear shape and to focal variation in section thickness (waviness), but (in relatively thick more than in very thick sections) the height of the peak of highest ploidy was falsely shortened, often markedly, and peak definition was worse. CONCLUSION: Choice of section thickness and selection of only center-containing nuclear sections for analysis with a bias in favor of elliptical nuclear section profiles in ultrathin sections are very important for optimal results; the choice of correction algorithm is less important.