この研究には、いいえユーザー施設契約の下で科学の DOE のオフィスである高度な光ソースのリソースが使用されます。デ-AC02-05CH11231。また夫妻・ l ・ Dobrzhinetskaya とモアッサ ナイト サンプルを貢献するため E. オバノン、スティワート オリーブ カタツムリ シェル データのオリーブのカタツムリの殻を準備するため H. 沈と周と教授 K. 陳オリーブのカタツムリの EDS 測定にしたいと思いますシェル。

<ol>
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K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synchrotron+X-ray+Micro-diffraction+-+Probing+Stress+State+in+Encapsulated+Thin+Silicon+Solar+Cells.">Synchrotron X-ray Micro-diffraction - Probing Stress State in Encapsulated Thin Silicon Solar Cells.</a> Procedia Engineering. 139, 123-133 (2016).</li><li>Xu, C. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elemental+Topological+Dirac+Semimetal:+&#945;-Sn+on+InSb(111)+.">Elemental Topological Dirac Semimetal: &#945;-Sn on InSb(111) .</a> Phys Rev Lett. 118 (14), 146402 (2017).</li><li>Chen, X., Tamura, N., MacDowell, A., James, R. 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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+high+stress+in+quartz+from+the+impact+site+of+Vredefort,+South+Africa.">Evidence for high stress in quartz from the impact site of Vredefort, South Africa.</a> Eur J Mineral. 23 (2), 169-178 (2011).</li><li>Jackson, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Material+and+Elastic+Properties+of+Al-Tobermorite+in+Ancient+Roman+Seawater+Concrete.">Material and Elastic Properties of Al-Tobermorite in Ancient Roman Seawater Concrete.</a> J Am Ceram Soc. 96 (8), 2598-2606 (2013).</li><li>Jackson, M. 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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parrotfish+Teeth:+Stiff+Biominerals+Whose+Microstructure+Makes+Them+Tough+and+Abrasion-Resistant+To+Bite+Stony+Corals.">Parrotfish Teeth: Stiff Biominerals Whose Microstructure Makes Them Tough and Abrasion-Resistant To Bite Stony Corals.</a> ACS Nano. 11 (12), 11856-11865 (2017).</li><li>Kunz, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dedicated+superbend+x-ray+microdiffraction+beamline+for+materials,+geo-,+and+environmental+sciences+at+the+advanced+light+source.">A dedicated superbend x-ray microdiffraction beamline for materials, geo-, and environmental sciences at the advanced light source.</a> Rev Sci Instrum. 80 (3), 035108 (2009).</li><li>Beckhoff, B., Kanngie&#223;er, B., Langhoff, N., Wedell, R., Wolff, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Practical X-Ray Fluorescence Analysis. , (2007).</li><li>Tamura, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=XMAS:+A+Versatile+Tool+for+Analyzing+Synchrotron+X-ray+Microdiffraction+Data.">XMAS: A Versatile Tool for Analyzing Synchrotron X-ray Microdiffraction Data.</a> Strain and Dislocation Gradients from Diffraction. , 125-155 (2014).</li><li>Dobrzhinetskaya, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moissanite+(SiC)+with+metal-silicide+and+silicon+inclusions+from+tuff+of+Israel:+Raman+spectroscopy+and+electron+microscope+studies.">Moissanite (SiC) with metal-silicide and silicon inclusions from tuff of Israel: Raman spectroscopy and electron microscope studies.</a> Lithos. , (2017).</li><li>Thibault, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+and+structural+crystallography+and+optical+properties+of+silicon+carbide+(SiC):+Part+II:+Structural+crystallography+and+optical+properties.">Morphological and structural crystallography and optical properties of silicon carbide (SiC): Part II: Structural crystallography and optical properties.</a> American Mineralogist. 29 (9-10), 327-362 (1944).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electron Backscatter Diffraction in Materials Science. , (2009).</li></ol>

著者が明らかに何もありません。

ALS のビームライン 12.3.2 複合 x 線回折と結晶試料の蛍光 x 線分析法を提案する.ラウエ回折、粉末回折も蛍光 x 線中自身が新しい手法、ビームライン 12.3.2 ミクロン スケール x 線ビーム径、検出器露出トリガー、および包括的に関連付けられているスキャン ステージ システムと同様に、それらを組み合わせた実験実験室の器械の不可能を可能にする解析ソフトウェアです。ビームライン光量?...

結晶構造解析はよくミクロン スケールで不均一性を表示します。地球科学研究科鉱物、結晶構造、および二次元系での位相関係の同定は物理学と化学の特定のシステムの両方を理解することが重要で、空間分解、定量的手法が必要です。たとえば、ローカライズされた 2D リージョン内位相分布に基づく鉱物間の関係を調べることができます。これは歴史と岩本体内で発生した化学的相互作用のための含意を持つことができます。また、単一の鉱物物質の構造を調べることができます。これは、鉱物であるかもしれないまたは現在 (ダイヤモンド ・ アンビル ・ セルなどのデバイスとその場で変形実験の場合など) にさらされている変形の種類を定めることができます。地球科学研究科エネルギーや波長分散型 x 線分光 (E/WDS) 電子後方散乱回折 (EBSD) と走査電子顕微鏡 (SEM) の組み合わせを使用してこれらの分析は、しばしば実行されます。ただし、試料調製が難しい、豊富な研磨、真空測定用マウントを含むできます。また、EBSD は常に隆起、侵食、または圧縮を経験していることが地質材料のための言い分ではない比較的無歪の結晶を必要とする表面技術です。
空間分解特性の 2次元 x 線回折および蛍光 x 線マッピングを使用しては、ALS のビームライン 12.3.2 のスケールで結晶の大きさがある単一または多相システムの広域マップを作るの迅速かつ簡単な方法は、数百ミクロンに (多結晶試料) の場合数ナノメートル。このメソッドには、他の一般的に使用される手法と比較して多くの利点があります。EBSD など、その他の 2次元結晶マッピング技術とは異なり回折サンプル周囲条件下で測定することができます、したがって真空チャンバーがないので特別な準備を必要は。回折が厳しい緊張またはプラスチック変形を経験しているものだけでなく、自然のままの結晶に適しています。試料の薄片、エポキシ樹脂に埋め込まれた材料としては一般的検討したも、岩や穀物を変更されず。データ コレクションは、高速、ラウエ回折のため通常よりも小さい 0.5 s/ピクセル未満の粉末回折のため 1 分/ピクセルと蛍光 x 線の 0.1 未満 s/ピクセルです。一時的にローカル ストレージと永続的をダウンロードする簡単だ、国立エネルギー研究科学的コンピューティング (ソルバー) センターでデータをローカルに保存します。ローカル クラスターまたは 20 分未満でソルバー クラスター、回折のデータ処理を実行できます。これにより短いデータの収集と分析、高速スループットと大面積測定の実験器具と比較した場合の期間です。
3 D プリントからすべてを分析する工学金属1,2, 太陽電池パネル変形に3ひずみとこのメソッドはさまざまなアプリケーション、特に材料科学で広く使用されていますトポロジカル物質4、記憶合金相に5、ナノ結晶材料6、7の高圧の動作に移行します。地球科学の最近のプロジェクトは、様々 な石英サンプル8,9火山セメント プロセス10,11, 方解石や霰石などミネラルのひずみの解析貝殻とサンゴの12,13または歯14、および隕石位相分布、新しい鉱物のミネラル構造同定に関する追加研究に含まれるアパタイトと高圧で塑性変形応答シリカも収集されています。ビームライン 12.3.2 で使用される技術はサンプル、鉱物学や岩石学的地域社会で誰にも関連するの広い範囲に適用されます。ここで我々 はビームライン 12.3.2、および地球科学分野で複合蛍光 x 線分析と粉/ラウエ回折法の有用性を実証するために現在のいくつかのアプリケーション用のデータ集録および解析プロトコルを概説します。
実験の詳細に入る前に話し合うエンド ステーションのセットアップに密接不離な関係だ (Kunzらの図 1および図 4を参照15). x 線ビーム蓄積リングを終了し、目的は再び焦点を当てる実験ハッチの入り口にソース、トロイダル ミラー (M201) が使用されます。セカンダリ ソース ポイントとしてロール スリットのセットの関数が通過します。それは (またはない) を単色、実験の種類に応じてスリットの 2 番目のセットを通過する前に、カークパ トリック バエズ (KB) ミラーのセットでミクロン サイズに焦点を当てています。ビームは、その信号を使用してビーム強度を決定、電離箱を通過します。イオン室に添付、ピンホール検出器に衝突から散乱信号をブロックすることです。集束ビームはサンプルを検出しました。サンプルは 8 モーターから成っているステージの上に置かれる: ラフ (下) x、y、z モータ、罰金 (上側) の x、y、z モータの 1 セットおよび 2 つの回転モーター (Φ および χ) の 1 つのセット。3 つの光学カメラで視覚化することができます: 低のズーム、高倍率ズームと電離箱の上部に配置され、1 つは、平面に配置した、x 線ビームと第二高ズーム カメラに関して約 45 ° の角度は、t に対して 90 ° の角度で配置彼はビーム x 線します。この最後の 1 つが縦向きのサンプル最高の作品 (例えば伝送モードの実験)、ピンホールに接続されているくさび型ミラーを用いたイメージングの実行と。X 線回折による検出器は大型回転ステージに位置し、角度と検出器の垂直変位の両方を制御できます。蛍光 x 線分析を収集するシリコンド リフト検出器はまたあります。サンプルの露出領域 (ROI) がフラット (ミクロン スケール) に限り、任意の方法で準備、発見または以上 50 〜 100 で覆われている μ m ポリイミド テープなど x 線透過材料の。
以下の手順では、反射のジオメトリで行われる、z 方向は通常のサンプルと x と y は、それぞれ水平方向と垂直方向の走査方向を想定しています実験について説明します。ステージと検出器システムの柔軟性のためしかし、いくつか実験を行い伝送ジオメトリで、x および z 方向は水平および垂直のスキャン方向 y が直接に平行ビーム (ジャクソンを参照してくださいら10,11)。

<table><tbody><tr><td>ThorLabs KB3x3 kinematic base, top half</td><td>ThorLabs</td><td>KBT3X3</td><td>Several of these bases are available for borrowing. The base must be the imperial and not the metric type, otherwise it will not properly fit on the stage.</td></tr><tr><td>Scotch double sided tape</td><td></td><td></td><td>Available at any office supply store, and also at the beamline</td></tr><tr><td>Polyimide/Kapton tape</td><td>Dupont</td><td></td><td>Several widths are commercially available. Any width that is enough to cover the sample is fine.</td></tr><tr><td>Samples</td><td></td><td></td><td>Provided by user, site of interest should be polished if larger mapping is desired.</td></tr><tr><td>Software: XMAS</td><td></td><td></td><td>Downloadable here https://sites.google.com/a/lbl.gov/bl12-3-2/user-resources</td></tr><tr><td>Software: IDL 6.2</td><td>Harris Geospatial Solutions</td><td></td><td></td></tr><tr><td>X-ray Diffraction Detector</td><td>DECTRIS Pilatus 1M&amp;nbsp;</td><td></td><td>hybrid pixel array detector</td></tr><tr><td>Huber stage</td><td></td><td></td><td>stage for detector</td></tr><tr><td>Vortex silicon drift detector&amp;nbsp;</td><td></td><td></td><td>silicon drift detector</td></tr><tr><td>IgorPro v. 6.37</td><td></td><td></td><td>Plotting software</td></tr></tbody></table>

synchrotron x-ray microdiffraction and fluorescence imaging of mineral and rock samples

本報告では、取得、x 線 microfluorescence (μXRF)、およびラウエ処理の詳細な手順について述べるし、粉末回折二次元 (2 D) マップ ビームライン 12.3.2 の高度な光ソース (ALS)、ローレンス ・ バークレイ国立実験室。測定は 10% 未満任意のサンプルで実行できる cm × 10 cm × 5 cm、平らな露出面で。標準的な材料 (蛍光 x 線分析、および Si、石英、または Al2O3回折など結晶試料の元素基準) を用いた実験的幾何学を校正します。X 線マイクロビームの焦点位置にサンプルを配置、マップの各ピクセルが 1 つの測定、例えば、 1 つの蛍光 x 線スペクトルまたは 1 つの回折パターンに対応、ラスター スキャンが実行されます。クリスマスは、出力テキスト ファイルには、各行がピクセル位置に対応社内開発ソフトウェアを使用して、データが処理されます。モアッサ ナイトと、オリーブのカタツムリの殻から代表的なデータは、データ品質、収集、および分析と戦略を示すに表示されます。

ラウエ回折
自然モアッサ ナイト (SiC) サンプル18最近の測定と分析を行った。サンプルだったし、カットして投資収益率を公開する研磨エポキシ プラグに埋め込まれた凝灰岩の部分成っていた。3 モアッサ ナイトの粒は、光学顕微鏡とラマン分光 (図 1 a) を使用して識?...

Watch this Scientific Journal Video about Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples at JoVE.com

Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples

ビームライン セットアップ ラウエ (多色放射) または回折 (単色放射) を使用して単一の結晶または粉末試料の急速な二次元 x 線蛍光と x 線回折のマッピングを実行するためのものについて述べる。作成したマップは、ひずみ、方向、位相分布および塑性変形に関する情報を与えます。

・鉱物及び岩石試料の蛍光を放射光 x 線回折

In this report, we describe a detailed procedure for acquiring and processing&#160;x-ray microfluorescence (&#956;XRF), and Laue and powder ...

synchrotron-x-ray-microdiffraction-fluorescence-imaging-mineral-rock

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Electronic Structure of Atoms

SCIENTISTS IN ACTION

8.9K ビュー.  Lawrence Berkeley National Laboratory. ビームライン セットアップ ラウエ (多色放射) または回折 (単色放射) を使用して単一の結晶または粉末試料の急速な二次元 x 線蛍光と x 線回折のマッピングを実行するためのものについて述べる。作成したマップは、ひずみ、方向、位相分布および塑性変形に関する情報を与えます。

ビデオ: Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples

Synthesis and Microdiffraction at Extreme Pressures and Temperatures

High pressure compounds and polymorphs are investigated for a broad range of purposes such as determine structures and processes of deep planetary interiors, design materials with novel properties, understand the mechanical behavior of materials exposed to very high stresses as in explosions or impacts. Synthesis and structural analysis of materials at extreme conditions of pressure and temperature entails remarkable technical challenges. In the laser heated diamond anvil cell (LH-DAC), very high pressure is generated between the tips of two opposing diamond anvils forced against each other; focused infrared laser beams, shined through the diamonds, allow to reach very high temperatures on samples absorbing the laser radiation. When the LH-DAC is installed in a synchrotron beamline that provides extremely brilliant x-ray radiation, the structure of materials under extreme conditions can be probed in situ. LH-DAC samples, although very small, can show highly variable grain size, phase and chemical composition. In order to obtain the high resolution structural analysis and the most comprehensive characterization of a sample, we collect diffraction data in 2D grids and combine powder, single crystal and multigrain diffraction techniques. Representative results obtained in the synthesis of a new iron oxide, Fe4O5 1 will be shown.

The laser heated diamond anvil cell combined with synchrotron micro-diffraction techniques allows researchers to explore the nature and properties of new phases of matter at extreme pressure and temperature (PT) conditions. Heterogeneous samples can be characterized in situ under high pressure by 2D mapping and combined powder, single-crystal and multigrain diffraction approaches.

エクストリーム圧力と温度での合成とMicrodiffraction

High pressure compounds and polymorphs are investigated for a broad range of purposes such as determine structures and processes of deep planetary ...

工学

Preparing Adherent Cells for X-ray Fluorescence Imaging by Chemical Fixation

X-ray fluorescence imaging allows us to non-destructively measure the spatial distribution and concentration of multiple elements simultaneously over large or small sample areas. It has been applied in many areas of science, including materials science, geoscience, studying works of cultural heritage, and in chemical biology. In the case of chemical biology, for example, visualizing the metal distributions within cells allows us to study both naturally-occurring metal ions in the cells, as well as exogenously-introduced metals such as drugs and nanoparticles. Due to the fully hydrated nature of nearly all biological samples, cryo-fixation followed by imaging under cryogenic temperature represents the ideal imaging modality currently available. However, under the circumstances that such a combination is not easily accessible or practical, aldehyde based chemical fixation remains useful and sometimes inevitable. This article describes in as much detail as possible in the preparation of adherent mammalian cells by chemical fixation for X-ray fluorescent imaging.

Here, we present a protocol on how to determine the quantity and distribution of metals in a sample using synchrotron X-ray fluorescence. We focus on adherent cells, and describe the chemical fixation method to prepare this sample. We then describe how to mount and image the sample using synchrotron X-rays.

化学固定することによりX線蛍光イメージングのための接着細胞の準備

X-ray fluorescence imaging allows us to non-destructively measure the spatial distribution and concentration of multiple elements simultaneously over ...

化学

Microcrystallography of Protein Crystals and In Cellulo Diffraction

The advent of high-quality microfocus beamlines at many synchrotron facilities has permitted the routine analysis of crystals smaller than 10 &#181;m in their largest dimension, which used to represent a challenge. We present two alternative workflows for the structure determination of protein microcrystals by X-ray crystallography with a particular focus on crystals grown in vivo. The microcrystals are either extracted from cells by sonication and purified by differential centrifugation, or analyzed in cellulo after cell sorting by flow cytometry of crystal-containing cells. Optionally, purified crystals or crystal-containing cells are soaked in heavy atom solutions for experimental phasing. These samples are then prepared for diffraction experiments in a similar way by application onto a micromesh support and flash cooling in liquid nitrogen. We briefly describe and compare serial diffraction experiments of isolated microcrystals and crystal-containing cells using a microfocus synchrotron beamline to produce datasets suitable for phasing, model building and refinement.
These workflows are exemplified with crystals of the Bombyx mori cypovirus 1 (BmCPV1) polyhedrin produced by infection of insect cells with a recombinant baculovirus. In this case study, in cellulo analysis is more efficient than analysis of purified crystals and yields a structure in ~8 days from expression to refinement.

Microcrystallography of Protein Crystals and In Cellulo Diffraction

A protocol is presented for X-ray crystallography using protein microcrystals. Two examples analyzing in vivo-grown microcrystals after purification or in cellulo are compared.

タンパク質結晶の微結晶化および<em&gt;セルロで</em&gt;回折

The advent of high-quality microfocus beamlines at many synchrotron facilities has permitted the routine analysis of crystals smaller than 10 &#181;m in ...

生化学

Dynamic Pore-scale Reservoir-condition Imaging of Reaction in Carbonates Using Synchrotron Fast Tomography

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve geologic seals and allow CO2 to escape. However, the dissolution processes at reservoir conditions are poorly understood. Thus, time-resolved experiments are needed to observe and predict the nature and rate of dissolution at the pore scale. Synchrotron fast tomography is a method of taking high-resolution time-resolved images of complex pore structures much more quickly than traditional &#181;-CT. The Diamond Lightsource Pink Beam was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 hours. The images were segmented and the porosity and permeability were measured using image analysis and network extraction. Porosity increased uniformly along the length of the sample; however, the rate of increase of both porosity and permeability slowed at later times.

Synchrotron fast tomography was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 h.

シンクロトロン高速断層撮影法を使用した炭酸における反応の動的細孔スケール貯水池条件イメージング

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve ...

Stress Distribution During Cold Compression of Rocks and Mineral Aggregates Using Synchrotron-based X-Ray Diffraction

We report detailed procedures for performing compression experiments on rocks and mineral aggregates within a multi-anvil deformation apparatus (D-DIA) coupled with synchrotron X-radiation. A cube-shaped sample assembly is prepared and compressed, at room temperature, by a set of four X-ray transparent sintered diamond anvils and two tungsten carbide anvils, in the lateral and the vertical planes, respectively. All six anvils are housed within a 250-ton hydraulic press and driven inward simultaneously by two wedged guide blocks. A horizontal energy dispersive X-ray beam is projected through and diffracted by the sample assembly. The beam is commonly in the mode of either white or monochromatic X-ray. In the case of white X-ray, the diffracted X-rays are detected by a solid-state detector array that collects the resulting energy dispersive diffraction pattern. In the case of monochromatic X-ray, the diffracted pattern is recorded using a two-dimensional (2-D) detector, such as an imaging plate or a charge-coupled device (CCD) detector. The 2-D diffraction patterns are analyzed to derive lattice spacings. The elastic strains of the sample are derived from the atomic lattice spacing within grains. The stress is then calculated using the predetermined elastic modulus and the elastic strain. Furthermore, the stress distribution in two-dimensions allow for understanding how stress is distributed in different orientations. In addition, a scintillator in the X-ray path yields a visible light image of the sample environment, which allows for the precise measurement of sample length changes during the experiment, yielding a direct measurement of volume strain on the sample. This type of experiment can quantify the stress distribution within geomaterials, which can ultimately shed light on the mechanism responsible for compaction. Such knowledge has the potential to significantly improve our understanding of key processes in rock mechanics, geotechnical engineering, mineral physics, and material science applications where compactive processes are important.

We report detailed procedures for compression experiments on rocks and mineral aggregates within a multi-anvil deformation apparatus coupled with synchrotron X-radiation. Such experiments allow quantification of the stress distribution within samples, that ultimately sheds light on compaction processes in geomaterials.

岩や放射光 x 線回折を用いた鉱物骨材の冷間圧縮時の応力分布

We report detailed procedures for performing compression experiments on rocks and mineral aggregates within a multi-anvil deformation apparatus (D-DIA) ...

環境学

Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography

This protocol describes fabricating microfluidic devices with low X-ray background optimized for goniometer based fixed target serial crystallography. The devices are patterned from epoxy glue using soft lithography and are suitable for in situ X-ray diffraction experiments at room temperature. The sample wells are lidded on both sides with polymeric polyimide foil windows that allow diffraction data collection with low X-ray background. This fabrication method is undemanding and inexpensive. After the sourcing of a SU-8 master wafer, all fabrication can be completed outside of a cleanroom in a typical research lab environment. The chip design and fabrication protocol utilize capillary valving to microfluidically split an aqueous reaction into defined nanoliter sized droplets. This loading mechanism avoids the sample loss from channel dead-volume and can easily be performed manually without using pumps or other equipment for fluid actuation. We describe how isolated nanoliter sized drops of protein solution can be monitored in situ by dynamic light scattering to control protein crystal nucleation and growth. After suitable crystals are grown, complete X-ray diffraction datasets can be collected using goniometer based in situ fixed target serial X-ray crystallography at room temperature. The protocol provides custom scripts to process diffraction datasets using a suite of software tools to solve and refine the protein crystal structure. This approach avoids the artefacts possibly induced during cryo-preservation or manual crystal handling in conventional crystallography experiments. We present and compare three protein structures that were solved using small crystals with dimensions of approximately 10-20 &#181;m grown in chip. By crystallizing and diffracting in situ, handling and hence mechanical disturbances of fragile crystals is minimized. The protocol details how to fabricate a custom X-ray transparent microfluidic chip suitable for in situ serial crystallography. As almost every crystal can be used for diffraction data collection, these microfluidic chips are a very efficient crystal delivery method.

Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography

This protocol describes in detail how to fabricate and operate microfluidic devices for X-ray diffraction data collection at room temperature. Additionally, it describes how to monitor protein crystallization by dynamic light scattering and how to process and analyze obtained diffraction data.

シリアル結晶の結晶 x 線回折その場その場での動的光散乱のためのマイクロ流体チップ

This protocol describes fabricating microfluidic devices with low X-ray background optimized for goniometer based fixed target serial crystallography. The ...

Sample Preparation and Transfer Protocol for In-Vacuum Long-Wavelength Crystallography on Beamline I23 at Diamond Light Source

Long-wavelength macromolecular crystallography (MX) exploits the anomalous scattering properties of elements, such as sulfur, phosphorus, potassium, chlorine, or calcium, that are often natively present in macromolecules. This enables the direct structure solution of proteins and nucleic acids via experimental phasing without the need of additional labelling. To eliminate the significant air absorption of X-rays in this wavelength regime, these experiments are performed in a vacuum environment. Beamline I23 at Diamond Light Source, UK, is the first synchrotron instrument of its kind, designed and optimized for MX experiments in the long wavelength range towards 5 &#197;. 
To make this possible, a large vacuum vessel encloses all endstation components of the sample environment. The necessity to maintain samples at cryogenic temperatures during storage and data collection in vacuum requires the use of thermally conductive sample holders. This facilitates efficient heat removal to ensure sample cooling to approximately 50 K. The current protocol describes the procedures used for sample preparation and transfer of samples into vacuum on beamline I23. Ensuring uniformity in practices and methods already established within the macromolecular crystallography community, sample cooling to liquid nitrogen temperature can be performed in any laboratory setting equipped with standard MX tools. 
Cryogenic storage and transport of samples only require standard commercially available equipment. Specialized equipment is required for the transfer of cryogenically cooled crystals from liquid nitrogen into the vacuum endstation. Bespoke sample handling tools and a dedicated Cryogenic Transfer System (CTS) have been developed in house. Diffraction data collected on samples prepared using this protocol show excellent merging statistics, indicating that the quality of samples is unaltered during the procedure. This opens unique opportunities for in-vacuum MX in a wavelength range beyond standard synchrotron beamlines.

Here, we present a protocol for cryogenic sample preparation and transfer of crystals into the vacuum endstation on beamline I23 at Diamond Light Source, for long-wavelength macromolecular X-ray crystallography experiments.

ダイヤモンド光源におけるビームラインI23上の真空中長波長結晶構造解析のためのサンプル調製および移送プロトコル

Long-wavelength macromolecular crystallography (MX) exploits the anomalous scattering properties of elements, such as sulfur, phosphorus, potassium, ...

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Imaging techniques are fundamental in order to understand cell organization and machinery in biological research and the related fields. Among these techniques, cryo soft X-ray tomography (SXT) allows imaging whole cryo-preserved cells in the water window X-ray energy range (284-543 eV), in which carbon structures have intrinsically higher absorption than water, allowing the 3D reconstruction of the linear absorption coefficient of the material contained in each voxel. Quantitative structural information at the level of whole cells up to 10 &#181;m thick is then achievable this way, with high throughput and spatial resolution down to 25-30 nm half-pitch. Cryo-SXT has proven itself relevant to current biomedical research, providing 3D information on cellular infection processes (virus, bacteria, or parasites), morphological changes due to diseases (such as recessive genetic diseases) and helping us understand drug action at the cellular level, or locating specific structures in the 3D cellular environment. In addition, by taking advantage of the tunable wavelength at synchrotron facilities, spectro-microscopy or its 3D counterpart, spectro-tomography, can also be used to image and quantify specific elements in the cell, such as calcium in biomineralization processes. Cryo-SXT provides complementary information to other biological imaging techniques such as electron microscopy, X-ray fluorescence or visible light fluorescence, and is generally used as a partner method for 2D or 3D correlative imaging at cryogenic conditions in order to link function, location, and morphology.

Here, a protocol describing the sample preparation and data collection steps required in cryo soft X-ray tomography (SXT) to image the ultrastructure of whole cryo-preserved cells at a resolution of 25 nm half pitch, is presented.

クライオ軟X線断層撮影による細胞の3D地図作成記述

Imaging techniques are fundamental in order to understand cell organization and machinery in biological research and the related fields. Among these ...

生物学

X-ray Fluorescence (XRF)

Source: Laboratory of Dr. Lydia Finney &#8212; Argonne National Laboratory
X-ray fluorescence is an induced, emitted radiation that can be used to generate spectroscopic information. X-ray fluorescence microscopy is a non-destructive imaging technique that uses the induced fluorescence emission of metals to identify and quantify their spatial distribution.

X-ray Fluorescence XRF: Elemental Analysis by Emitted Fluorescent X-Ray

蛍光 x 線 (XRF)

Source: Laboratory of Dr. Lydia Finney &#8212; Argonne National Laboratory
X-ray fluorescence is an induced, emitted radiation that can be used to ...

JoVE Science Education

分析化学

Single Crystal and Powder X-ray Diffraction

Source: Tamara M. Powers, Department of Chemistry, Texas A&#38;M University&#160;
X-ray crystallography is a technique that uses X-rays to study the structure of molecules. X-ray diffraction (XRD) experiments are routinely carried out with either single-crystal or powdered samples.
Single-crystal XRD:
Single-crystal XRD allows for absolute structure determination. With single-crystal XRD data, the exact atomic positions can be observed, and thus bond lengths and angles can be determined. This technique provides the structure within a single crystal, which does not necessarily represent the bulk of the material. Therefore, additional bulk characterization methods must be utilized to prove the identity and purity of a compound.
Powder XRD:
Unlike single-crystal XRD, powder XRD looks at a large sample of polycrystalline material and therefore is considered a bulk characterization technique. The powder pattern is considered a &#34;fingerprint&#34; for a given material; it provides information about the phase (polymorph) and crystallinity of the material. Typically, powder XRD is used to study minerals, zeolites, metal-organic frameworks (MOFs), and other extended solids. Powder XRD can also be used to establish bulk purity of molecular species.
Previously, we have seen how to grow X-ray quality crystals (see video in Essentials of Organic Chemistry series). Here we will learn the principles behind XRD. We will then collect both single-crystal and powder data on Mo2(ArNC(H)NAr)4, where Ar = p-MeOC6H5.

単結晶および粉末 x 線回折

Source: Tamara M. Powers, Department of Chemistry, Texas A&#38;M University&#160;
X-ray crystallography is a technique that uses X-rays to study the ...