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Plant building aggregates, components and parts of optical instruments

Plant building aggregates, components and parts of optical instruments

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VIDEO ON THE TOPIC: Optical Instruments

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Goal 9: Industry, Innovation and Infrastructure

Floating riverine aggregates are composed of a complex mixture of inorganic and organic components from their respective aquatic habitats. Their architecture and integrity are supplemented by the presence of extracellular polymeric substances of microbial origin.

They are also a habitat for virus-like particles, bacteria, archaea, fungi, algae, and protozoa. In this study we present different confocal laser scanning microscopy strategies to examine aggregates collected from the Danube and Elbe Rivers.

In order to collect multiple types of information, various approaches were necessary. Small aggregates were examined directly. To analyze large and dense aggregates, limitations of the technique were overcome by cryo-sectioning and poststaining of the samples. The staining procedure included positive staining specific glycoconjugates and cellular nucleic acid signals as well as negative staining aggregate volume and multichannel recording.

Data sets of cellular nucleic acid signals CNAS and the structure of aggregates were visualized and quantified using digital image analysis. The Danube and Elbe Rivers differed in their aggregate composition and in the relative contribution of specific glycoconjugate and CNAS volume to the aggregate volume; these contributions also changed over time. We report different spatial patterns of CNAS inside riverine aggregates, depending on aggregate size and season.

Based on our samples, we discuss the strengths and challenges involved in scanning and quantifying riverine aggregates. In rivers, primary particles are frequently and perhaps characteristically transported as larger flocculated aggregates. They are structurally very stable because they are exposed to a constant shear force, resulting in relatively small aggregates compared to aggregates in lakes and marine systems 41 , Abiotic mechanisms such as physical coagulation, collision frequency, and stickiness are involved in particle aggregation These aggregates, which may be regarded as mobile biofilms e.

Apart from water, biofilms may consist of dissolved, colloidal, and particulate materials varying in size and composition They are composed of a complex mixture of components including inorganic minerals , living organic bacteria, archaea, fungi, algae, protozoa, and viruses , and nonliving organic extracellular polymeric substances [EPS], allochthonous and autochthonous detritus, lignins, tannins, etc.

Cellular material within a biofilm can vary greatly. The actual structure of the biofilm matrix varies greatly depending on the microbial cells present, their physiological status, the nutrients available, and the prevailing physical conditions Knowledge about the structure and the function of aggregates, both in environmental and engineered systems, is very important In engineered systems such as wastewater treatment plants, understanding flocculation can help in the management of that process.

In environmental systems, such structure-function relationships can provide ecologically relevant information about material transfers between particulate and dissolved matter or about spatial distribution of microorganisms, with the related impacts on the aquatic food web.

Numerous methods are available to help characterize aggregate properties. Microscopic as well as photographic techniques have been used to analyze aggregate structure. In recent years, confocal laser scanning microscopy CLSM data sets have allowed the visualization and quantification of three-dimensional 3-D structures 18 , In this study, we analyzed aggregates from the Danube and Elbe Rivers by collecting reflection, nucleic acid, glycoconjugate, and negative stain signals using CLSM.

In order to receive multifarious information about the aggregates, various approaches were necessary: small aggregates were examined directly, and large and dense aggregates were physically sectioned and poststained. Although most of the detected nucleic acid signals derive from bacteria, we refer to them as cellular nucleic acid signals CNAS including potential archaea and virus signals. Nucleic acid signals can potentially also be obtained from fungi, algae, and protozoa.

But the detection of extracellular DNA can be excluded due to its type of appearance 7. Data sets of specific glycoconjugates, CNAS, and aggregate structure were visualized and quantified by using digital image analysis. The distribution of CNAS within riverine aggregates was determined by autocorrelation.

Based on our samples, we describe the strengths and challenges in scanning and quantifying riverine aggregates using CLSM. Additionally, we discuss structure, function, and potentially important differences in aquatic aggregates from these two large European rivers. Both locations are in the braided river region and, therefore, can be regarded as representative and comparable areas. We sampled repeatedly during an annual period to accommodate typical seasonal changes in water discharge, temperature, and variable suspended solid loads and quality.

Hence, water samples from the Danube River were taken in spring 3 May , summer 20 July , fall 13 October , and winter 8 January From the Elbe River, water samples were taken in spring 21 March , summer 8 August , fall 3 November , and winter 23 January Sampling depth was approximately 30 cm.

Riverine aggregates were sampled in 1-liter Plexiglas bottles. With an inverted ml glass pipette 14 , the aggregates were carefully transferred to Eppendorf tubes, where staining was performed.

Chemical analyses of nitrate, nitrite, ammonium, orthophosphate, total soluble reactive phosphorus, and total phosphorus were performed based on German Standard Methods for the Examination of Water, Wastewater and Sludge. Determinations of total suspended solids, particulate organic matter, and chlorophyll- a concentrations are published elsewhere The lectins were self-labeled according to the data sheet of the supplier.

The EPS glycoconjugates were stained with one of the lectins as described previously Briefly, for lectin staining, the lectins were diluted with deionized water to a final concentration of 0. One hundred microliters of this solution was added to each sample and incubated for 20 min in the dark at room temperature.

The aggregates were then carefully washed three times with tap water to remove unbound lectins and were never allowed to dry in the air.

SYBR Green I is a nucleic acid-specific stain widely used to detect prokaryotes and virus particles in aquatic environments The dimensions and volumes of the aggregates were determined by a negative staining procedure. Lawrence et al. We took advantage of this method to determine aggregate volumes. Different probes and fluorochrome-staining solutions such as dextrans Invitrogen , rhodamines Invitrogen , fluorescein Sigma Aldrich, St. These stains and probes are characterized by their chemical properties and molecular weights MWs and were used at different concentrations.

Probes and fluorochrome-staining solutions such as dextrans, rhodamines, fluorescein, and Cy5 were not appropriate for this analysis, probably due to their MWs or their binding properties data not shown. Negative staining with Alexa Fluor allowed us to calculate the aggregate volumes. Hence, to analyze our samples, we stained the water phase—the volume of the scanned box which was not occupied by the aggregate—with the fluorochrome Alexa Fluor C 37 H 33 N 3 O 13 S 2 ; MW of The positively stained samples were carefully transferred into Cover Well imaging chambers 0.

The aggregate structure was analyzed by CLSM using visible lasers nm, nm, and nm. Emission signals were detected from to nm inorganic and mineral compounds; detected by reflection signals , to nm DNA signals; detected with SYBR Green I , to nm negative staining signals; detected with Alexa Fluor , and from to nm lectin signals; detected with fluorochrome Cy5 or Alexa Fluor Conventional analyses of larger riverine aggregates traditional staining procedure and scanning with a view from the top were limited by laser penetration and diffusion of staining solutions.

To obtain information on the distribution of lectin-specific glycoconjugates and CNAS inside large riverine aggregates, cryo-sections were performed To analyze cryo-sections with the software ImageJ, special plug-ins were developed.

After thresholds were established, the binary image of each cryo-section was divided into three sectors. Within each sector, one subsample area of rectangular shape extending across the whole dimension of the aggregate's cryo-section was chosen see Fig. The dimension of the aggregate was given by the lectin-specific glycoconjugate. The software cropped each subsample, and pixels of the CNAS were quantified. Single scans of different cryo-sections showing distribution of CNAS including potential archaea and virus signals inside an aggregate a and d.

Insets in panels a and d show glycoconjugate and CNAS. The white rectangle indicates a subsample for CNAS volume calculation. The distribution of the calculated CNAS volumes within the subsamples of the cryo-sections is presented b and e. Spatial autocorrelation shows the distribution and accumulations of CNAS within the cryo-sections of the aggregates c and f. The first local maximum provides information on CNAS layer width. Secondary maxima indicate a regular spatial distribution of CNAS within the cryo-section, pointing to the repeating occurrence of CNAS layers whose width can be estimated from the first maximum.

Black lines indicate the level of significance twice the standard error. Color allocation: green, nucleic acid; red, glycoconjugates; yellow, CNAS in or in contact with lectin-specific extracellular polymeric substances. This program can handle multichannel data and colocalization of data.

For each channel and data set, the threshold was set manually. Due to the very heterogeneous composition of the aggregates, automatic batch processing could not be applied. To visualize 3-D data sets, Imaris, version 4. For each channel and image, thresholds were set manually. Adobe Photoshop CS2 was used to insert calibration bars into the images. For statistical analysis, the software program SPSS, version The criteria of normal distribution Kolmogorov-Smirnov test and homogeneity of variances Levene test were not met, so seasonal differences in the data were determined with a nonparametric test Friedman test.

A Tamhane posthoc test which does not require homogeneity of variances in the data was used to determine significant differences between seasons.

Differences between the two rivers were tested using a Mann-Whitney U test. To determine the distribution of CNAS within riverine aggregates, cryo-sections were analyzed. Only those subsamples that included a minimum of 10 slices containing CNAS were included in the analysis. Two approaches were used: the first was used to detect abundance differences between the subsamples of the three sectors, and the second was used to characterize the internal structure of the cryo-sections using a finer spatial resolution.

The mean abundances of CNAS within each subsample were calculated. Then, differences between the two outer subsamples and the middle subsample were computed to detect potential spatial accumulation of cells. The averages of these two differences were tested against zero using a Wilcoxon test. Mean differences between abundances will only be significantly different from zero if CNAS are more abundant either on the two outer subsamples or in the middle subsample of the cryo-section.

To test whether CNAS occurred in accumulations anywhere within the cryo-sections of the aggregates and to test for regularity in distributions of CNAS within the cryo-sections, we calculated the spatial autocorrelation If the autocorrelation values were higher than twice the standard error for positive autocorrelations, they were considered significant.

The highest autocorrelations always occurred at lag 1 and then typically decreased uniformly. The first local maximum was defined as the number of lags with significant positive autocorrelations, starting from lag 1 until the significance level was reached or the autocorrelations started to increase again. This first maximum provides information on the width of the CNAS layers.

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Aaron Toponce

Floating riverine aggregates are composed of a complex mixture of inorganic and organic components from their respective aquatic habitats. Their architecture and integrity are supplemented by the presence of extracellular polymeric substances of microbial origin. They are also a habitat for virus-like particles, bacteria, archaea, fungi, algae, and protozoa. In this study we present different confocal laser scanning microscopy strategies to examine aggregates collected from the Danube and Elbe Rivers.

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Или надумает продать кольцо. Беккер не мог ждать. Он решительно поднял трубку, снова набрал номер и прислонился к стене. Послышались гудки.

Беккер разглядывал зал.

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В голове у него не было ни единой мысли - полная пустота. Он не знал ни где он находится, ни кто его преследует и мчался, подгоняемый инстинктом самосохранения. Он не чувствовал никакой боли - один лишь страх. Пуля ударила в кафельную плитку азульехо чуть сзади.

Aaron Toponce

Мидж Милкен явно чего-то не поняла. - Это многое объясняет, - настаивала.  - Например, почему он провел там всю ночь.

- Тебе удалось стереть электронную почту Хейла. - Нет, - сконфуженно ответила .

- Мне показалось, что я уловил в вашей речи бургосский акцент. Сам я из Валенсии. Что привело вас в Севилью. - Я торговец ювелирными изделиями.

How to Calculate Overall Equipment Effectiveness: A Practical Guide

У нас тут творятся довольно странные вещи. Я хотел спросить… - Черт тебя дери, Джабба! - воскликнула Мидж.  - Именно это я и пыталась тебе втолковать.

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Goal 9: Industry, Innovation and Infrastructure

Он понимал, что времени у него. Агенты могут появиться в любую минуту. Собрав все силы, Хейл, сильнее обхватив Сьюзан за талию, начал пятясь подниматься по лестнице. Она пыталась цепляться каблуками за ступеньки, чтобы помешать ему, но все было бесполезно.

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