An Application of the Frozen Sectioning Technic for Cutting Serial Sections Thru the Brain

The availability of CO₂ ice makes it practical to cut large blocks of cerebral tissue by the freezing method. If the tissue is first treated with 20-30% ethyl alcohol for sufficient time to secure uniform penetration of the alcohol (about 24 hours), formation of hard ice crystals can be controlled and serial sections 25-100 μ thick can be cut with negligible loss. The alcohol can be added to the fixative used for perfusion, or it can be added at any time later in the firing process, or after fixation is completed. The sections are cemented to the slide and groups of slides are manipulated thru staining processes in glass trays. Ordinary cell and fiber stains give satisfactory results. The method is particularly useful for certain neurophysiological purposes such as defining the location of electrode tracks and lesions and certain types of retrogrades. The Prussian blue test for electrolytically deposited iron can be conveniently applied in conjunction with other stains, to determine the point at which a given action potential response was observed, if steel electrodes are used.     

眼结膜印迹细胞法及其影响因素

本文摸索眼结膜印迹细胞法及其影响因素,证明该方法是一项简便,快速,有效地评价机体维生素Ａ营养状况的方法;取样材料,染色时间,温度,透明方法均可影响制片结果。该方法适合基层推广应用。     

Microdistribution of benthic invertebrates in a rocky mountain (U.S.A.) stream

A study of the benthic invertebrate community inhabiting a small, foothill trout stream in the Rocky Mountains of Idaho was conducted over a two-year period. Monthly Hess samples and short-term experiments using substratum-filled trays were used to describe the spatial dispersion of the benthos and to examine the response of invertebrate populations to substratum and current. A method was devised for measuring available surface area which involved coating individual stones with latex and measuring the area of the print resulting from inking the impression left on the latex mold.The dispersion of all populations was clumped throughout the year. Alteration of the cross-sectional pattern of current velocity and stream bed composition changed the pattern of distribution but not the extent of clumping. Collections made in areas of depositing and eroding substrata revealed a more diverse fauna in the latter. Most groups of organisms found in the riffle were scarcer in the pools or absent from them. The pool fauna contained no important additions over those found in the riffles.After a year's study of invertebrate populations in an otherwise undisturbed riffle, the substratum was altered and the flow made more uniform; an increase in the abundance of most of the benthic invertebrates followed. No single factor was responsible for the increase, but the change in substratum size and degree of compaction accounted for most of the change. Interpretation of the results was aided by findings from experiments using substratum-filled trays.Two series of stream experiments using the trays were conducted: one to test the relative importance of current and substratum and the other to test the effect of particle size on the distribution of the benthic fauna. In the first series, placement of trays of stones in a pool resulted in an increase in numbers of some but not all of the invertebrates over numbers usually occurring in the pool. Trays filled with stones and placed in a riffle supported fewer animals than found on the adjacent stream bed but more than in the pool. Variations are attributed to differences in current velocity and amounts of imported organic and inorganic debris. Three different relationships of population numbers to current velocity were found for different members of the community (direct, indirect, and parabolic) over the range of 10 to 60 cm/sec. The second series of experiments consisted of two sets of trays filled with stones of medium or large pebbles, respectively. Nine taxa, as well as all of the combined taxa, showed a preference for trays of small stones over the natural stream bed. A few taxa were noticeably more abundant on the small substratum than on the large but most of the fauna showed only slight increases in numbers or remained constant on the two substrata. Only three taxa showed a direct relation of numbers to total surface area presented by the stones.Number and kinds of organisms found in trays filled with a uniform size of substratum did not correspond to those taken in Hess samples from the natural stream bed. This has important implications in terms of currently recommended pollution monitoring techniques. However, it is suggested that if the substratum composition of the trays more nearly matched that of the stream, the correspondence would be much better. The results of the present study also throw considerable doubt on the adequacy of generalizations derived from earlier studies of responses to substratum size and suggest several reasons for reevaluating current ideas regarding the influence of substratum on invertebrate distribution.     

The effect of food supply on the colonisation of barley by aerially dispersing spiders (Araneae)

A field experiment was carried out to determine whether different levels of food availability affected the retention rate of ballooning spiders landing in trays of seedling barley plants, half of which were infested with aphids from laboratory cultures. The trays were placed within bases in the field, then collected sequentially and spider numbers assessed in each tray. Deposition trays, containing trapping fluid only, were used to measure ballooning activity throughout the experimental period. The experiment was repeated four times. Overall, ballooning spiders were more likely to be retained in trays where aphid prey were present, with a total of 340 spiders found in the infested trays and 251 in the aphid-free trays, over the four experiments. Most of the spiders found were of the family Linyphiidae. In the second and fourth experiments the increased retention of spiders in the aphid-infested trays was statistically significant. Immature linyphiids alone also showed significantly higher retention in the infested trays in those two experiments and in the fourth experiment were largely responsible for the higher numbers found in the infested trays. There was also a statistically significant trend for a higher retention rate of female spiders, compared to males, in the barley trays than would have been expected from the ratios of females to males caught in the deposition trapping trays alone.     

Use of oviposition trays to estimate numbers of wheat bulb fly Leptohylemyia coarctata eggs

To obtain reliable estimates of wheat bulb fly populations in time to provide advice on seed treatment of the following crop a method of collecting eggs in trays of soil placed in fields has been devised. The trays are examined by 20 August when half of the eggs have been laid.     

A simple practical method for bulk and rapid extraction of free-living nematodes from marine and estuarine sediments

A simple, rapid method is described for the extraction of large numbers of free-living nematodes from estuarine sediments. This method does not physically or chemically alter or damage the nematodes, but instead relies on their downward movement through a filtering layer of double ply tissue paper and into aerated water-filled trays. Seven trials each with 10 trays kept at 25?°C for an initial period of 24 h yielded 3985 live nematodes l^?1 (± 511.5 standard deviation) of estuarine sediment, free of sediment and with minimal debris. Time effects were statistically significantly different, with the same 10 trays yielding another 1259 nematodes l^?1 (± 413.4) when kept for a second period of 24 h at the same temperature. Temperature effects were also significant, and 7 trials each with 10 trays kept for 24 h at 20–21?°C, produced a lower yield of 2160 nematodes l^?1 (± 532.7) of sediment. The method is expected to be of use in nematode extractions from both estuarine and marine sediments.     

Foraging of multimammate mice, Mastomys natalensis, under different predation pressure: cover, patch-dependent decisions and density-dependent GUDs

Patch use under predation risk often results in a change of feeding behaviour in the prey animals. However, such changes only appear if the animals are able to assess under which predation pressure they live. We investigated patch use of Mastomys natalensis under different conditions of avian predation pressure.
In replicated maize field plots in Morogoro, Tanzania, avian predators were allowed under natural conditions (control), attracted with perches and nest boxes or kept out with nets. During four one‐week periods in late 1999, we measured rodent feeding decisions with the giving‐up density (GUD) method. Trays with known amounts of millet seeds in sand were placed in pairs, one of them under a cover, the other one in the open. M. natalensis mice were expected to give up sooner in the open trays than in those with cover. We hypothesised that M. natalensis mice could assess the ambient predation pressure leading to larger difference in GUD between covered and non‐covered trays in the plots where predators were attracted. We also made video recordings of the rodent activity at a pair of trays in each treatment. The GUD‐values were significantly lower for the covered trays but predation pressure did not affect this difference. The video observations showed that in the control and netted plots the animals visited trays equally frequently regardless of the cover, while the visits in the predator‐attracted plots occurred significantly more often in the covered trays. We conclude that M. natalensis can assess the ambient predation pressure and adapt its behaviour at a feeding patch. However, the variation in predation pressure in our experiment was not obvious from the GUD. Moreover, we found a strong relation between rodent density and GUD, which may mask variations in perceived predation pressure. Similar GUD values may be reached in different ways and we present models to investigate whether animals’ decision to forage at a food patch is only affected by the seed density at that patch, not by that at a neighbour patch.     

Workability and productivity of robotic plug transplanting workcell

Summary Transplanting is a necessary operation in transplant production systems. Transplanting operation is labor-intensive and automation can reduce labor costs. Plugs are actively growing young transplants with two well-defined morphologic parts: the stem-leaf portion and the root-growth medium portion. They may be grown in regularly situated cells on traylike containers. This regularity makes plugs suitable for automated transplanting operations. It is, therefore, beneficial for in vitro plant propagation systems to include plugs as intermediate products before they are delivered to the greenhouses. Flexible automation and robotics technologies have been applied to develop a robotic workcell for transplanting plugs from plug trays to growing flats. Main components of the workcell include a robot, an end-effector, and two conveyer belts for transporting trays and flats. The end-effector for extracting, holding, and planting plugs is a “sliding-needles-with-sensor” gripper. The sensor signals the robot to complete a transplanting cycle only when a plug is properly held by the gripper. Systems analysis and computer simulation were conducted to study factors affecting workability and productivity of various workcell designs. These factors included: dimensions and kinematics of the robot and its peripheral equipment, layout and materials flow, fullness of plug trays, and successful extraction rate of plugs. The analysis also indicated that machine vision systems could add valuable capabilities to the workcell, such as robot guidance and plug quality evaluation. Engineering economic analysis was performed to investigate the interaction of workcell technical feasibility and economic viability. Presented in the Session-in-Depth Robotics in Tissue Culture at the 1991 World Congress on Cell and Tissue Culture, Anaheim, California, June 16–20, 1991.     

Method for analyzing outlines with an application to recent Japanese crania

If the visual image and quantitative treatment of a form could be merged, the combination could become a powerful analytical method in morphology. A method is presented for creating an averaged outline. In this method, since the averaged outline is produced statistically, hypothesis testing can be performed on the visual image of the outlines. Intra- and intergroup variation of the outlines can then be assessed by taking allometry into account. This method is applied to the analysis of sexual differences in the midsagittal outline of recent (1880s-1920s) Japanese crania. The averaged outlines of both sexes showed subtle features that could not be detected by traditional measuring techniques. Because the roundness, flatness, or protrusion of specific aspects of the morphology are clearly apparent, sexual differences in this sample could be identified. Changes resulting from allometry did not seem to be present in either sex. The shape distance (DIR) introduced here matched well the visual impression derived from comparisons of the Japanese sample with a Jomon (Neolithic) male and the Broken Hill specimen.     

A single serum dilution method for the quantitation of neutralizing antibodies to varicella-zoster virus

A one-point serum dilution method for determination of neutralizing antibody in human sera to varicella-zoster (V-Z) virus instead of the serial serum dilution method was investigated. Focus counting was performed under a microscope on day 5 to 6 after inoculation of V-Z virus into 6-well plastic trays in which human embryonic lung cells were grown. A table was constructed to estimate the ND50 titers by the per cent reduction of the focus count from the control at only one dilution of test sera. The estimated ND50 values agreed well with those determined by the serial serum dilution method. Test sera showed a slight nonspecific reactivity at low serum dilutions, but reliable results could usually be obtained at a serum dilution of 1:8 or more. This method, which saves materials and labor, was applied to the quantification of neutralizing antibody against V-Z virus in human sera with satisfactory accuracy and reproducibility.     

Comparative analysis of different impression techniques in relation to single tooth impression

It is of interest to compare the accuracy of three different impression techniques for a single tooth impression. We used 3 groups with 15 samples each in this study. Group 1: Putty and light body in a sectional stock tray; Group 2: Monophase and extra light body in a sectional stock tray; Group 3: Matrix impression technique. 15 impressions were taken of a prepared tooth on a typodont with each technique. The dimensions of the casts poured from these impression techniques were compared with the control typodont tooth. Data analysis shows that the matrix impression technique gave the best results in terms of dimensional study followed by monophase and extra light body impression technique and putty and light body impression technique gave the least accurate results. The results show that there is a statistically significant difference between the three impression techniques in terms of dimensional stability. Data analysis shows that the matrix impression technique gave the best results in terms of dimensional study followed by monophase and extra light body impression technique and putty and light body impression technique gave the least accurate results. The variations between the groups are within acceptable limits. Hence, it can be concluded that all the impression techniques will result in adequate dimensional stability and can be used in clinical scenarios.     

Evaluation of a testing method for the fatigue performance of total knee tibial trays.

Catastrophic failure of tibial baseplates due to fatigue fracture have occurred in patients. The International Standards Organisation (ISO) have proposed an endurance test to ensure a safety level for trays with respect to this problem. At present the magnitude of the applied load has not been defined. The purpose of this study is to evaluate the ISO test by comparing the results of two tibial trays: one with a clinical history of fracture of the tibial baseplate in some cases (Kinematic) and one without such a record (Kinemax). A load of 2000 N was selected, representing the peak force during the normal walking cycle. Finite element analysis (FEA) predicted that both the Kinematic and Kinemax trays would rapidly fail for two defined contact positions. Mechanical testing performed on both these components confirmed the FEA predictions. However, under service conditions, the Kinematic tray has been reported to have a failure of only 1-2%, the failure occurring at a mean of 54 months for early fracture and 7 years for late fracture. There are no published reports of failed Kinemax trays. It is concluded that the ISO test is an exaggeration of reality when a physiological load is used. As a result of our study we propose that a load of 500 N would effectively screen components with respect to the ISO test conditions for the likelihood of fracture.     

A convenient microtitre tray procedure for yeast identification

Media for yeast identification tests were incorporated into the wells of a microtitre tray. The tests included fermentation and assimilation of carbohydrates, assimilation of nitrogen compounds, growth in vitamin-free medium, resistance to cycloheximide, and observations for cell morphology and sporulation. Results of tests conducted in the trays showed very good agreement with those obtained by conventional methods. Eighteen reference yeasts were correctly identified from tests conducted in the trays. The trays of media could be stored, and provided a convenient system for yeast identification.     

The effect of natural enemies on the spread of barley yellow dwarf virus (BYDV) by Rhopalosiphum padi (Hemiptera: Aphididae)

The effects of two natural aphid enemies, adult Coccinella septempunctata Linneaus, a predator, and Aphidius rhopalosiphi de Stefani Perez, a parasitoid, on spread of barley yellow dwarf virus (BYDV) transmitted by the bird cherry-oat aphid, Rhopalosiphum padi (Linnaeus) were studied under laboratory conditions. Predators or parasitoids were introduced to trays of durum wheat seedlings and the patterns of virus infection were observed after two, seven and 14 days of exposure. More plants were infected with BYDV in control trays without A. rhopalosiphi than in trays with the parasitoid present, both seven and 14 days after the introduction of parasitoids. Patterns of virus infection were found to be similar over time in trays with a parasitoid present and in control trays. More plants were infected in trays with C. septempunctata present than in control trays, both two and seven days after the introduction of the coccinellid. The spread of virus infections progressed differently over time for the two treatments (predator and parasitoid), differences between treatments being most marked after two days and seven days, when more plants exposed to predators but fewer exposed to parasitoids were infected with BYDV compared to their respective controls. However, by the 14th day 88% of all plants were infected and there was no significant difference between the two treatments. The role of natural enemies in spread of BYDV is discussed.     

A convenient microtitre tray procedure for yeast identification

Media for yeast identification tests were incorporated into the wells of a microtitre tray. The tests included fermentation and assimilation of carbohydrates, assimilation of nitrogen compounds, growth in vitamin-free medium, resistance to cycloheximide, and observations for cell morphology and sporulation. Results of tests conducted in the trays showed very good agreement with those obtained by conventional methods. Eighteen reference yeasts were correctly identified from tests conducted in the trays. The trays of media could be stored, and provided a convenient system for yeast identification.     

A behavioral method for separation of house fly (Diptera: Muscidae) larvae from processed pig manure

A behavioral method applicable in biodegradation facilities for separation of house fly (Musca domestica L.) larvae from processed pig manure is presented. The method is based on placing a cover over the larval rearing tray, while escaping larvae are collected in collection trays. Separation units must be placed in a dark room to avoid negative phototactic responses of the larvae. After 24 h of separation, over 70% of the larvae escaped from processed manure and were collected in collection trays. Most of the larvae pupated within 48 h after separation. Mean weight of pupae recovered from manure residue was not significantly different from mean weight of pupae of separated individuals. Eclosion rate of pupae recovered from manure residue was significantly lower than eclosion of separated individuals, and was strongly related to separation success. Factors responsible for escape behavior of larvae are discussed.     

Experimental colonization of sand, gravel and stones by macroinvertebrates in the Acheron River, southeastern Australia

SUMMARY. 1. The contribution of drift to macroinvertebrate colonization was studied over 10 days using pairs of substratum-filled trays, one suspended above the streambed (and thus able to be colonized by drift only) and one buried flush with the streambed (and thus open to colonization from all directions). Trays were filled with sand, gravel or stones.
2. Colonization of sand-filled trays was rapid, being completed within 24h (i.e. no subsequent change in numbers of individuals per tray); colonization of gravel or stone-filled trays was not completed by the end of the experiment.
3. Drift contributed an average of 86% of the colonizing fauna of the sand-filled trays, 36% of the fauna of the gravel-filled trays and 25% of the fauna of the stone-filled trays. The low contribution of the drift to the gravel and stones is at odds with some studies which, using similar techniques, suggest that drift is the primary source of colonizing individuals.
4. Most individual species showed low contributions by drift to colonization, with the majority having contributions under 66%.
5. Distinct differences were noted between the fauna colonizing the sand-filled trays and those colonizing the gravel or stone-filled trays (which were similar).
6. Several species common in the drift were rare in the colonizing fauna and several species which were common colonizers were absent or rare in the drift.     

A rapid screening technique for lymphocytotoxic antibodies using tray-frozen lymphocytes

A method whereby sera can be screened for the presence of lymphocytotoxic antibodies within 4 hr using lymphocytes frozen in microtest trays is described. The reactions of the sera of 48 hemodialysis patients against freshly prepared lymphocytes were compared with those against tube-frozen (384 reaction pairs) and tray-frozen (864 reaction pairs) cells. There was a better than 90% concordance, and only 3% of the reactions differed from negative to strongly positive or vice versa. Preliminary results indicate that the method is also suitable for B-cell (HLA-DR) antibody testing. Routine screening against 50- or 100-cell panels can be accomplished more rapidly and more efficiently using tray-frozen lymphocytes. The data matrix for analysis of a serum's HLA specificity is greatly reduced in comparison to the conventional screening technique, rendering the method particularly suitable for microcomputer processing.     

Auditory imagery associated with Japanese onomatopoeic representation

When we want to express the subjective impression of sound, we usually use onomatopoeias. We investigated the relationship between the impression of auditory imagery associated with Japanese onomatopoeic representation and onomatopoeic features. Subjects rated the impressions of auditory imageries associated with 40 onomatopoeias on semantic differential scales. Principal component analysis was applied to the rating data and the scales were integrated to the components of beauty, potency and sharpness. We determined the relationship between phonetic features of onomatopoeias and tonal features of the auditory imageries on the obtained principal components. Onomatopoeic representations that included voiced consonants were associated with a "dirty" impression. Onomatopoeic representations that contained approximant sounds were associated with a "powerful" impression. The vowel /i/ was associated with a "sharp" impression and vowels /u/ and /o/ were associated with a "dull" impression. The obtained tendencies confirmed that some onomatopoeic features reflected particular impressions of auditory imagery.     

Rearing of larvae of Anopheles stephensi, using water replacement, purification and automated feeding

For rearing progenies of single females of Anopheles stephensi (Liston) (Diptera: Culicidae), a method is proposed using small trays with perforated side walls (called open trays) to allow for water replacement and purification using an aquatic plant Vallisneria spiralis. The performance of this method has been compared with that of rearing single progenies in trays without mesh side walls (closed trays). Effects of larval density and food quantity on rearing performance have been studied in both types of trays. In open trays the larvae developed about 2 days faster than in closed trays. Male larvae developed significantly faster (0.4 days) than females, but this difference was not significantly influenced by method of rearing or larval density. Adults from fast developing larvae were heavier than those emerging from slowly developing larvae from a single egg batch. Open trays gave higher larval survival than closed trays. A system has been developed to automatize larval feeding.

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Elevage des larves issues de femelles isolées d' Anopheles stephensi Liston (Dipt.; Culicidae), avec utilisation d'un système automatique d'alimentation et de renouvellement de l'eau

Résumé Une technique, utilisant des récipients avec des parois latérales perforées (baptisés récipients ouverts) permettant le renouvellement de l'eau et sa purification par une plante aquatique (Vallisneria piralis), est préconisée pour élever la descendance de femelles isolées de A. stephensi. Les performances de cette technique ont été comparées à l'élevage dans des récipients sans parois perforées (récipients clos). L'influence de la densité et de la quantité d'aliments a été examinée dans les deux cas. Dans les récipients ouverts les larves se développent plus vite (deux jours de moins); les mâles se développent plus vite que les femelles, mais cette différence (0,4 j) n'est significiativement pas influencée par les techniques d'élevage ou la densité. Pour une même ponte les adultes provenant des larves se développant le plus vite sont plus gros que ceux qui sont issus des larves les plus lentes. Le récipient ouvert donne une meilleure survie. Un système a été mis au point pour automatiser l'alimentation.