Data reported in this work are part of the USDA-ARS National Program 306 Project 'Values-Added Products from Cottonseed' research supported by the Agency's in-house funding. Publication of this paper is supported in part by the Journal of Visualized Experiments. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer. We acknowledge the constructive comments from JoVE science editor and peer reviewers in the review and revision process.

1.綿実油と大豆ミールベースの​​製品（図1） <ol><li>市販の供給源から、原材料、脱脂綿実油と大豆の食事を取得します。 </li><li> 0.5ミリメートルの鋼スクリーン16を通過させるようにサイクロンサンプルミルで固形脱脂食事を粉砕することにより作業の食事を取得します。 </li><li>食事21中の水溶性成分を分離する：水抽出（200ミリリットル水25グラムの食事）後に作業し、食事からの水で洗浄食事を準備。 </li><li>タンパク質はアルカリ抽出し、酸沈殿16で作業し、食事から隔離する準備します。 </li></ol>ウッド単板ストリップの調製<ol><li>ストリップ長い88.9ミリメートル、幅25.4ミリメートルに市販のソースから取得したカット木材ベニア（1.59ミリメートルの厚さ）。 </li><li>鉛筆は、各ストリップの一端から長さ25.4ミリメートル（1.0 &quot;）で木目を越え行をマーク適切にテストする治療法または数字でこれらのストリップにラベルを付け5。。 -10木材対は、各テストの変数が用意されている。 </li></ol>接着剤スラリーの調製<ol><li>適用率（ 例えば、4 mgの乾燥含有センチ-2）により、テスト用の木材標本ごとに必要な水洗浄し、食事の量を計算し、総接着面積（ 例えば、2.54 X 2.54センチメートル接合面積の90木材片の581センチメートル×2各）プラスenoughness約30％の追加（ すなわち、水の4×581のx 130％3g）を、例えば、食事をした。 </li><li>脱イオン水（3：25 w / w）の水で洗浄食事を混合し、パラフィルムで密封し、ビーカー中で2時間、磁気撹拌棒で撹拌する。 </li></ol>集成材試験片の4準備<ol><li> 25.4ミリメートル（1.0 &quot;）の長さをカバーする2木材ベニヤストリップの一端に接着剤スラリーを磨き空気乾燥10のために - 。15分または粘着性まで。 </li><li>再び第1層の上部と空気乾燥で接着剤スラリーの第二の層を磨く。乾燥した接着剤の量製剤が適用ボンディング1cm 2当たり乾燥固体約4.5mg、各木材片であるている。 </li><li> 2木材ベニヤストリップの粘着性の接着剤コーティングされた領域をオーバーラップ（25.4 X 25.4ミリメートルまたは1.0 &quot;×1.0&quot;）。 400 PSI（2.8メガパスカル）の圧力で20分間、100℃でベンチトップ加熱プレスを使用したホットプレス。圧力は木材サンプルの重なり面積で割っプレスによって加えられた力であることに注意してください。各試験の変数に必要に応じてこれらの接合のパラメータを変更してもよい。 </li><li>クール空調室や湿度制御（22の温度- 23℃、相対湿度50 - 60％; 図2）インキュベーター中で48時間集成試料を調整。 </li></ol> 5.水の抵抗実験<ol><li> （ - 23°C 22）、室温でプラスチックトレーで48時間水道水に、初期状態調節した後、結合した木材標本を浸し。浸漬後の湿った標本は、せん断のためにすぐにテストされている破断強度と湿潤強度として報告した。ベニヤの表面上の過剰の水は穏やかに測定前にティッシュペーパーで軽く叩くことにより除去することができる。 </li><li> 18 O / N（室内条件（60％ - - 23℃、相対湿度50 22の温度）で乾燥、次いで、4時間63ºCの水浴中で、最初のコンディショニングの後、結合した木材片の別のセットを浸し - 20時間）。 48時間の乾燥時間で一回浸漬乾燥サイクルを繰り返します。乾燥した試験片は、その後破断時のせん断強度について試験し、浸した接着強度として報告されている。 </li></ol> 6.ラップせん断強度の測定<ol><li> 7メガパスカルの把持圧力で材料試験機上の32×40ミリメートル耐つまとびグリッドウェッジグリップに結合した木材片を取り付け、そして分-1 1ミリメートルで、クロスヘッド速度を設定。 </li><li>各結合した木材標本のための破断時のせん断強度を測定し、記録します。複数の測定の結果は、各接着剤形のために平均化されるulationとテスト変数。 </li></ol>

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The authors have nothing to disclose.

本稿では、木材接着剤として植物種子ベースの製品を準備し、テストするための基本的な手順を提示します。このプロトコルでexampled接着剤スラリーは単に脱脂粕製品および水である。種々の接着剤配合物（例えば、ドデシル硫酸ナトリウム、亜硫酸水素ナトリウムまたはキリ油など）の試験試薬（例えば、pH、固体と水の比）混合条件で5,6,23および/ ​​または変更3,24を添?...

木材の接着は、森林製品産業の増加役割を果たしており、効率的な木材資源1を利用するための重要な要因である。木材のための天然物ベースの接着剤の使用への関心はこの期間の後、石油系接着剤の価格は、彼らがいくつかの伝統的な市場からのタンパク質接着剤を避難ほど低くなった。1960年2付近にピークに到達するために、1930年代から着実に増加した。過去20年間では、この傾向は、再生可能な生分解性、そしてより環境に許容される材料の使用における新たな関心が逆しています。これらの天然資源は、綿実タンパク質6、米ぬか7、小麦グルテン8、蒸留穀物タンパク質9、キャノーラタンパク質および油10-12、ソルガムからリグニン及びサトウキビバガス13が挙げられるが、これらに限定されるものではないが、大豆タンパク質3-5 、14、エビの殻15から導出多糖類。 &lt;p個のクラスは= &quot;jove_content&quot;&gt;種子タンパク質単離物は、広く潜在木材接着剤として評価されているのに対し、単離手順は、腐食アルカリ性および酸性の試薬 ​​を含み、それは、比較的高価少ない環境に優しい16の接着剤ベースを分離することができます。このように、治療の有無にかかわらず、いくつかの脱脂シードの食事（粉）も、タンパク質が17〜19を隔離するように、これらの食事の粘着特性が同様に機能しないにもかかわらず、接着剤の目的のためにテストされています。私たちは、順次異なる画分に綿実粕（CM）を分画し、接合木材ベニヤ20,21での接着強度を調べた。水不溶性固形分（以下、洗浄綿実粕、WCM）は綿実油タンパク質単離物（CSPI）に匹敵する、木材接着剤として使用することができ、CSPIも調製することが低コストになる。 接着強度および耐水性の性能を評価する2つの重要なパラメータである潜在的な接着材料。ここで、接着強さは、各木材片のラップ結合の破断時の剪断強度として報告される。接着剤の耐水性は、水浸漬の破断時の集成試料の重ね剪断強度の変化によって測定される。原料として脱脂綿実油と大豆の食事を使用して、このプロトコルは、木材接着剤として、テスト植物種子ベースの製品を製造するためのシンプルで簡単な方法を提供します。このプロトコルは、天然物ベースの木材接着剤の多くの経済的、環境に配慮した製剤を求める中で努力を促進する上で参考になる。

<table border="0" cellpadding="0" cellspacing="0" width="927">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Material and Equipment</td>
			<td style="width:227px;">Supplier/Manufacturer</td>
			<td style="width:227px;"></td>
			<td style="width:211px;">Address</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;">defatted cottonseed meal</td>
			<td>Kentwood Co-op</td>
			<td></td>
			<td>Kentwood, LA, USA&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">defatted soy meal</td>
			<td>Kentwood Co-op</td>
			<td></td>
			<td>Kentwood, LA, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">wood veneers</td>
			<td>Certainly Wood, Inc.</td>
			<td></td>
			<td>East Aurora, NY, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cyclone sample mill (model 3010-014)</td>
			<td>UDY Corporation</td>
			<td></td>
			<td>Fort Collins, CO, USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Benchtop Heated Press&#160; (Model 3856)</td>
			<td>Carver, Inc.</td>
			<td></td>
			<td>Wabash, IN. USA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Materials Tester</td>
			<td>Zwick GmbH &#38; Co.</td>
			<td></td>
			<td>Ulm, Germany</td>
		</tr>
	</tbody>
</table>

preparation and testing of plant seed meal-based wood adhesives

これらの植物原料を再生可能な、環境に優しいと考えられているように最近では、木材接着剤などの植物種子ミールベースの​​製品への関心が着実に増加している。これらの天然物は、環境持続可能性の懸念を緩和するために、石油系接着剤の代替物として機能することができる。この作品は、原料として綿実油および大豆ミールを用いて植物種子ベースの木材接着剤の準備とテストを示しています。未処理の食事に加えて、水は食事を洗浄し、タンパク質単離物を調製し、試験する。接着剤スラリーを2時間（w / wの3時25）、脱イオン水で凍結乾燥ミール製品を混合することによって調製される。各貼付製剤は、ブラシを使用して2木材ベニアストリップの一端に印加される。粘着性の接着剤は、木材ベニヤストリップはホットプレスによってラップと接着されているの領域を被覆した。接着強度は、破断時の接合木材片のせん断強度として報告されている。接着剤の耐水性により測定される集成材のせん断強度の変化は、水浸漬後にブレークで標本。このプロトコルは1つが、合成ベースの木材接着剤の置換のための適切な候補として植物種子ベースの農産物を評価することができます。または添加剤や接着条件なしの接着剤配合の調整は、様々な実用的なアプリケーションのために彼らの接着特性を最適化することができます。

各接着剤配合物の性能は、破断時の接合木材片のせん断強度によって決定された値が使用される木材単板の寸法に応じて変化する。で推奨綿実油-2のより厚い、より広いストリップとは対照的に、例えば、 表1に、接合試験片の乾燥および浸漬し、接着強度の値は、（綿実油-1を参照）より薄く、狭いメープルストリップを使用する場合に低い同じ綿実油系接着剤製剤を使用して?...

Watch this Scientific Journal Video about Preparation and Testing of Plant Seed Meal-based Wood Adhesives at JoVE.com

Preparation and Testing of Plant Seed Meal-based Wood Adhesives

To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

植物種子ミールベースの​​木材接着剤の調製および試験

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable ...

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8.6K ビュー.  United States Department of Agriculture. To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

ビデオ: Preparation and Testing of Plant Seed Meal-based Wood Adhesives

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

トウモロコシとテオシンテ線における ウスチラゴ の病原性を評価するための迅速かつ効率的な方法

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

環境学

Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the exploration of renewable resources are necessary to overcome these challenges. Within the multinational framework MultiBioPro we are developing biorefinery pipelines to maximize the use of plant biomass. More specifically, we use poplar and tobacco tree (Nicotiana glauca) as target crop species for improving saccharification, isoprenoid, long chain hydrocarbon contents, fiber quality, and suberin and lignin contents. The methods used to obtain these outputs include GC-MS, LC-MS and RNA sequencing platforms. The metabolite pipelines are well established tools to generate these types of data, but also have the limitations in that only well characterized metabolites can be used. The deep sequencing will allow us to include all transcripts present during the developmental stages of the tobacco tree leaf, but has to be mapped back to the sequence of Nicotiana tabacum. With these set-ups, we aim at a basic understanding for underlying processes and at establishing an industrial framework to exploit the outcomes. In a more long term perspective, we believe that data generated here will provide means for a sustainable biorefinery process using poplar and tobacco tree as raw material. To date the basal level of metabolites in the samples have been analyzed and the protocols utilized are provided in this article.

Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline.

バイオベースの産業の原料としてたばこ木とポプラの評価のための転写産物および代謝物プロファイリング

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the ...

A Technical Perspective in Modern Tree-ring Research - How to Overcome Dendroecological and Wood Anatomical Challenges

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to environmental changes and to finally reconstruct such changes. This is done by analyzing growth variations back in time and correlating various plant-specific parameters to (for example) temperature records. Integrating wood anatomical parameters in these analyses would strengthen reconstructions, even down to intra-annual resolution. We therefore present a protocol on how to sample, prepare, and analyze wooden specimen for common macroscopic analyses, but also for subsequent microscopic analyses. Furthermore we introduce a potential solution for analyzing digital images generated from common small and large specimens to support time-series analyses. The protocol presents the basic steps as they currently can be used. Beyond this, there is an ongoing need for the improvement of existing techniques, and development of new techniques, to record and quantify past and ongoing environmental processes. Traditional wood anatomical research needs to be expanded to include ecological information to this field of research. This would support dendro-scientists who intend to analyze new parameters and develop new methodologies to understand the short and long term effects of specific environmental factors on the anatomy of woody plants.

Here we present a protocol outlining how to sample wooden specimens for the overall assessment of their growth structures. Macro- and microscopic preparation and visualization techniques necessary to generate well-replicated and highly resolved wood anatomical and dendroecological dataset, are described are described.

現代の木リング研究の技術的な観点 -  Dendroecologicalと木材解剖課題を克服する方法

Dendroecological research uses information stored in tree rings to understand how single trees and even entire forest ecosystems responded to ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

毒性のための飲料水試料の迅速な評価のためのRTgill-W1細胞とのインピーダンスベースの流体バイオチップの製造および試験

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

フィールド条件の下で種子発芽、休眠、および死亡率を評価するための信頼性の高い方法

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

On the Preparation and Testing of Fuel Cell Catalysts Using the Thin Film Rotating Disk Electrode Method

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a high surface area carbon, and to test their performance in thin film rotating disk electrode (TF-RDE) measurements. The TF-RDE methodology is widely used for catalyst screening; nevertheless, the measured performance sometimes considerably differs among research groups. These uncertainties impede the advancement of new catalyst materials and, consequently, several authors discussed possible best practice methods and the importance of benchmarking.
The visual tutorial highlights possible pitfalls in the TF-RDE testing of Pt/C catalysts. A synthesis and testing protocol to assess standard Pt/C catalysts is introduced that can be used together with polycrystalline Pt disks as benchmark catalysts. In particular, this study highlights how the properties of the catalyst film on the glassy carbon (GC) electrode influence the measured performance in TF-RDE testing. To obtain thin, homogeneous catalyst films, not only the catalyst preparation, but also the ink deposition and drying procedures are essential. It is demonstrated that an adjustment of the ink&#39;s pH might be necessary, and how simple control measurements can be used to check film quality. Once reproducible TF-RDE measurements are obtained, determining the Pt loading on the catalyst support (expressed as Pt wt%) and the electrochemical surface area is necessary to normalize the determined reaction rates to either surface area or Pt mass. For the surface area determination, so-called CO stripping, or the determination of the hydrogen underpotential deposition (Hupd) charge, are standard. For the determination of the Pt loading, a straightforward and cheap procedure using digestion in aqua regia with subsequent conversion of Pt(IV) to Pt(II) and UV-vis measurements is introduced.

Preparing and testing Pt/C fuel cell catalysts is subject to continuous discussion in the scientific community with respect to reproducibility and best practice. With the presented work, we intend to present a step-by-step tutorial to make and test Pt/C catalysts, which can serve as benchmark for novel catalyst systems.

準備および薄膜回転ディスク電極法を用いた燃料電池触媒のテストに

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

植物感染試験: スプレーと傷を介した植物病原体イネいもち病菌接種

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

Xylem Water Distribution in Woody Plants Visualized with a Cryo-scanning Electron Microscope

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water distribution in plant tissues in combination with freeze fixation techniques using liquid nitrogen (LN2). For woody species, however, preparations for observing the xylem transverse-cut surface involve some difficulties due to the orientation of wood fibers. Additionally, higher tension in the water column in xylem conduits can occasionally cause artifactual changes in water distribution, especially during sample fixation and collection. In this study, we demonstrate an efficient procedure to observe the water distribution within the xylem of woody plants in situ&#160;by using a cryostat and cryo-SEM. At first, during sample collection, measuring the xylem water potential should determine whether high tension is present in the xylem conduits. When the xylem water potential is low (&#60; ca. &#8722;0.5 MPa), a tension relaxation procedure is needed to facilitate better preservation of the water status in xylem conduits during sample freeze fixation. Next, a watertight collar is attached around the tree stem and filled with LN2&#160;for freeze fixation of the water status of xylem. After harvesting, care should be taken to ensure that the sample is preserved frozen while completing the procedures of sample preparation for observation. A cryostat is employed to clearly expose the xylem transverse-cut surface. In cryo-SEM observations, time adjustment for freeze-etching is required to remove frost dust and accentuate the edge of the cell walls on the viewing surface. Our results demonstrate the applicability of cryo-SEM techniques for the observation of water distribution within xylem at cellular and subcellular levels. The combination of cryo-SEM with non-destructive in situ observation techniques will profoundly improve the exploration of woody plant water flow dynamics.

Observing the water distribution within the xylem provides significant information regarding water flow dynamics in woody plants. In this study, we demonstrate the practical approach to observe xylem water distribution in situ&#160;by using a cryostat and cryo-SEM, which eliminates artifactual changes in the water status during sample preparation.

クライオ走査型電子顕微鏡で可視化された木質植物のキシレム水分布

A scanning electron microscope installed cryo-unit (cryo-SEM) allows specimen observation at subzero temperatures and has been used for exploring water ...

Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of the woodboring beetle, Chlorophorus caragana Xie &#38; Wang (2012), for both types of electron microscopy. The scanning electron microscopy (SEM) sample preparation protocol was based on sample chemical fixation, dehydration in a series of ethanol baths, drying, and sputter-coating. By adding Tween 20 (Polyoxyethylene sorbitan laurate) to the fixative and the wash solution, the insect body surface of woodboring beetle was washed more cleanly in SEM. This study&#39;s transmission electron microscopy (TEM) sample preparation involved a series of steps including fixation, ethanol dehydration, embedding in resin, positioning using fluorescence microscopy, sectioning, and staining. Fixative with Tween 20 enabled penetrate the insect body wall of woodboring beetle more easily than it would had been without Tween 20, and subsequently better fixed tissues and organs in the body, thus yielded clear transmission electron microscope observations of insect sensilla ultrastructures. The next step of this preparation was determining the positions of insect sensilla in the sample embedded in the resin block by using fluorescence microscopy to increase the precision of target sensilla positioning. This improved slicing accuracy.

In order to observe ultrastructure of insect sensilla, scanning and transmission electron microscopy (SEM and TEM, respectively) sample preparation protocol were presented in the study. Tween 20 was added into the fixative to avoid sample deformation in SEM. Fluorescence microscopy was helpful for improving slicing accuracy in TEM.

木管化カブトムシの付属物の走査型および透過型電子顕微鏡の試料調製方法

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of ...

The Effect of Construction and Demolition Waste Plastic Fractions on Wood-Polymer Composite Properties

Construction and demolition waste (CDW), including valuable materials such as plastics, have a remarkable influence on the waste sector. In order for plastic materials to be re-utilized, they must be identified and separated according to their polymer composition. In this study, the identification of these materials was performed using near-infrared spectroscopy (NIR), which identified material based on their physical-chemical properties. Advantages of the NIR method are a low environmental impact and rapid measurement (within a few seconds) in the spectral range of 1600-2400 nm without special sample preparation. Limitations include its inability to analyze dark materials. The identified polymers were utilized as a component for wood-polymer composite (WPC) that consists of a polymer matrix, low cost fillers, and additives. The components were first compounded with an agglomeration apparatus, followed by production by extrusion. In the agglomeration process, the aim was to compound all materials to produce uniformly distributed and granulated materials as pellets. During the agglomeration process, the polymer (matrix) was melted and fillers and other additives were then mixed into the melted polymer, being ready for the extrusion process. In the extrusion method, heat and shear forces were applied to a material within the barrel of a conical counter-rotating twin-screw type extruder, which reduces the risk of burning the materials and lower shear mixing. The heated and sheared mixture was then conveyed through a die to give the product the desired shape. The above-described protocol proved the potential for re-utilization of CDW materials. Functional properties must be verified according to the standardized tests, such as flexural, tensile, and impact strength tests for the material.

Secondary material streams have been shown to include potential raw materials for production. Presented here is a protocol in which CDW-plastic waste as a raw material is identified, followed by various processing steps (agglomeration, extrusion). As a result, a composite material was produced, and mechanical properties were analyzed.

木高分子複合物に及ぼす構造・解体廃プラスチック分率の影響

Construction and demolition waste (CDW), including valuable materials such as plastics, have a remarkable influence on the waste sector. In order for ...

植物種子ミールベースの​​木材接着剤の調製および試験

植物種子ミールベースの木材接着剤の調製および試験