Name: in situ characterization of shewanella oneidensis mr1 biofilms by salvi and tof-sims
Uploaded: 2017-08-18T13:00:00
Description: Watch this Scientific Journal Video about In Situ Characterization of Shewanella oneidensis MR1 Biofilms by SALVI and ToF-SIMS at JoVE.com

太平洋の北西国立研究所 (なり) 地球と生物科学 (EBD) ミッション種監督研究と開発 (LDRD) 基金へのサポート感謝しております。インストゥルメンタル アクセスが w. r. ワイリー環境分子科学研究所 (EMSL) 一般ユーザーの提案書を提供されました。EMSL はオフィスの生物と環境研究 (BER) なりで主催国立科学ユーザー施設です。著者は博士 『 鼎の証拠、原稿を読んで、有用なフィードバックを提供することをありがちましょう。バテル契約 DE AC05 76RL01830 下 DOE のなりが運営しています。

1 材料準備 <ol><li>ミディアム チューブの準備 <ol><li>(バイオ フィルム文化と成長曲線ごとに必要な 3 つごとに必要な 1 つ) 血清ボトル 注: 前述の導入は、任意の成長で。このプロシージャの興味の細菌の緊張のために必要な栄養素を提供するために適切なメディアを利用できます。この場合、&#34; ナノワイヤー &#34; メディアとブドウ糖媒体なし TSB s. oneidensis 氏 1 GFP ?...

<ol>
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C., Wingender, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biofilm+matrix.">The biofilm matrix.</a> Nat Rev Microbiol. 8 (9), 623-633 (2010).</li><li>Aldeek, F., 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=Patterned+hydrophobic+domains+in+the+exopolymer+matrix+of+Shewanella+oneidensis+MR-1+biofilms.">Patterned hydrophobic domains in the exopolymer matrix of Shewanella oneidensis MR-1 biofilms.</a> Appl Environ Microbiol. 79 (4), 1400-1402 (2013).</li><li>Hua, X., 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=In+situ+molecular+imaging+of+a+hydrated+biofilm+in+a+microfluidic+reactor+by+ToF-SIMS.">In situ molecular imaging of a hydrated biofilm in a microfluidic reactor by ToF-SIMS.</a> Analyst. 139 (7), 1609-1613 (2014).</li><li>Yu, X. Y., Yang, L., Cowin, J. P., Iedema, M., Zhu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systems+and+methods+for+analyzing+liquids+under+vacuum.">Systems and methods for analyzing liquids under vacuum.</a> US Patent. , (2013).</li><li>Yu, X. Y., Liu, B., Yang, L., Zhu, Z., Marshall, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+electrochemical+device+and+process+for+chemical+imaging+and+electrochemical+analysis+at+the+electrode-liquid+interface+in+situ.">Microfluidic electrochemical device and process for chemical imaging and electrochemical analysis at the electrode-liquid interface in situ.</a> US Patent. , (2014).</li><li>Yang, L., Yu, X. Y., Zhu, Z. H., Thevuthasan, T., Cowin, J. 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X., Ma, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+Graphene-Modified+Electrodes+in+Microbial+Fuel+Cells.">Applications of Graphene-Modified Electrodes in Microbial Fuel Cells.</a> Materials. 9 (10), (2016).</li><li>Yu, J., Zhou, Y., Hua, X., Zhu, Z., Yu, X. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+Situ+Characterization+of+Hydrated+Proteins+in+Water+by+SALVI+and+ToF-SIMS.">In Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS.</a> J Vis Exp. (108), e53708 (2016).</li><li>Hill, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Effects of Electron-Transport-System Impairment on Hydrogen Gas Production by the Bacterium Shewanella oneidensis MR-1. , (2007).</li><li>McCormick, A. J., 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=Biophotovoltaics:+oxygenic+photosynthetic+organisms+in+the+world+of+bioelectrochemical+systems.">Biophotovoltaics: oxygenic photosynthetic organisms in the world of bioelectrochemical systems.</a> Energy Environ Sci. 8 (4), 1092-1109 (2015).</li><li>Liu, B., 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=In+situ+chemical+probing+of+the+electrode-electrolyte+interface+by+ToF-SIMS.">In situ chemical probing of the electrode-electrolyte interface by ToF-SIMS.</a> Lab Chip. 14, 855-859 (2014).</li><li>Keune, K., Hoogland, F., Boon, J. J., Peggie, D., Higgitt, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+"added+value"+of+SIMS:+A+mass+spectrometric+and+spectroscopic+study+of+an+unusual+Naples+yellow+oil+paint+reconstruction.">Evaluation of the "added value" of SIMS: A mass spectrometric and spectroscopic study of an unusual Naples yellow oil paint reconstruction.</a> Int J mass Spectrom. 284 (1-3), 22-34 (2009).</li><li>Lee, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mass Spectrometry Handbook. , 988 (2012).</li><li>Petrovic, M., Barcelo, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+anionic+and+nonionic+surfactants,+their+degradation+products,+and+endocrine-disrupting+compounds+in+sewage+sludge+by+liquid+chromatography/mass+spectrometry.">Determination of anionic and nonionic surfactants, their degradation products, and endocrine-disrupting compounds in sewage sludge by liquid chromatography/mass spectrometry.</a> Anal Chem. 72 (19), 4560-4567 (2000).</li><li>Vickerman, J. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tof+Sims+Study+of+the+Desorption+of+Emulsifiers+from+Polystyrene+Latexes.">Tof Sims Study of the Desorption of Emulsifiers from Polystyrene Latexes.</a> Surf Interface Anal. 21 (6-7), 387-394 (1994).</li><li>Pe&#241;uelas-Urquides, 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=Measuring+of+Mycobacterium+tuberculosis+crowth.+A+correlation+of+the+optical+measurements+with+colony+forming+units.">Measuring of Mycobacterium tuberculosis crowth. A correlation of the optical measurements with colony forming units.</a> Braz J Microbiol. 44 (1), 287-289 (2013).</li><li>Dohnalkova, A. 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ログ段階で接種後数日とバイオ フィルムが 3.1 の手順で説明するよう健康とイメージングのため十分な厚さだ前に拡大する温度をテストすることが重要です。S. oneidensis 室温; MR1 バイオ フィルムを養殖具体的手順します。しかし別の部屋の温度の成長率に影響を与えます。したがって、バイオ フィルムは TOF-SIMS 解析に進む前に準備ができているかどうかを理解する光イメージング技術を使?...

細菌バイオ フィルム細菌前記細胞が接続し、多くの可能な環境の中で生き残ることができる有害な物理的・機械的刺激の変化を生き残るためにための防衛として時間をかけて進化してきた表面関連のコミュニティであります。1,2バイオ フィルム大幅に調査した、生体臨床医学、医用生体工学、農業、産業の研究・開発など多くの分野のアプリケーションがあります。1,2セルフ プロデュースに細胞外高分子物質 (EPS) と、ローカル水クラスター環境などを含むこれらの複雑な微生物の化学マッピングを理解する正確を得る不可欠ですな彼らの生物学的活動の描写です。2
バイオ フィルムは存在し、高含水状態の内で育ちます。これは、揮発性液体の真空の勉強の難しさのため飛行時間二次イオン質量分析 (TOF-SIMS) など真空表面解析テクニックを使用して偉大な課題を提示します。その結果、真空ベースの表面分析技術は、乾燥状態だけでバイオ フィルム サンプルを勉強にほとんど専ら限られています。しかし、乾燥状態でバイオ フィルムを勉強して、真の生物学的微小環境の正確な調査を阻害します。それはしばしば、EPS の整合性とバイオ フィルム形態、その場で液体研究にバイオ フィルムの乾燥質量スペクトル結果を比較した後示されている抜本的な変更が発生します。3,4この記事分析液体真空インターフェイス (サルヴィ)、5,6マイクロ流体リアクターでの我々 のシステムの使用を採用することにより自然な水和状態内のバイオ フィルムを勉強するためのソリューションを提示します。したがって真空内液体マトリックスの構造の整合性を維持しながら二次イオンビーム プローブへの直接アクセスを提供するマイクロ ポリジメチルシロキサン (PMDS) の薄いシリコン窒化 (罪) 膜の下で液体が入っています。商工会議所。7,8
S. oneidensis氏 1 緑色蛍光タンパク質 (GFP) を表現する変異は、その代謝の多様性と環境、応用微生物学、基づいていたで一般的な使用のためこのバイオ フィルムの手順図のモデル生物として選ばれました。大きく上金属還元剤や細胞外の電子転送のための独自の機能。9,10,11また、GFP の存在は簡単連続バイオ フィルム厚蛍光顕微鏡による監視、フルオレセイン イソチオ シアン酸 (FITC) フィルターを使用しての許可。私たちの以前の研究では、最大 100 μ m の厚さにバイオ フィルムの成長のためoperando の蛍光イメージングを使用して罪ウィンドウに添付ファイルを好むこの細菌の証拠を示しています。4,12本稿が蛍光顕微鏡によるバイオ フィルムの存在の確認を説明するだけのサルヴィは超解像などその他の光学撮像方法と互換性のある蛍光イメージング (すなわち、構造化照明顕微鏡 (SIM)9) および共焦点レーザー走査顕微鏡 (CLSM) 画像4)。光イメージングは、バイオ フィルムの厚さを測定する機能し、それは、その厚さと罪ウィンドウへの付着を確認する表示されます、バイオ フィルムの形状の 3 D イメージを取得できます。9 GFP は、SIMS 分析で使用された、GFP なしS の oneidensisを成長曲線光学密度のこの唯一の必要な測定として使用され、任意の蛍光イメージングを必要としません。一般的に、タグ付きの GFP の違いと成長曲線のタグの付いていない種が重要ではありません。さらに、このプロトコルは、手順を記述するモデル生物としてs. oneidensis氏 1 GFP を使用して、この手順はサルヴィ内栽培のために必要とされるすべての菌株の設計されています。とはいえ、必要な細菌株の知識を与え、時間、温度、酸素環境など生育条件が使用される細菌の歪みに合わせて変更する必要があります。成長媒体のためこの手順は養殖で、ブドウ糖とブドウ糖なしトリプシン大豆寒天 (TSA) なし「ナノワイヤー」中, トリプシン大豆スープ (TSB) を使用します。成長のため、膜の拡張を監視するため、「ナノワイヤ」中の組成は、特別策定されていますと、細いワイヤと培地成分形を取るように見えるs. oneidensisのペリプラズムがされている以前の研究で確立。13,14
その場で当社の以前の議定書液体 TOF-SIMS サルヴィは、タンパク質の固定化と TOF-SIMS 解析およびデータ圧縮に関する詳細なプロトコルと同様に、罪に添付ファイルを提供している利点について説明しました。12データ削減の手順を繰り返しよりもむしろ本稿になる代わりにセットアップおよびバイオ フィルムの存在と事前の厚みを検出する画像処理の手順と同様に、私たちのサルヴィのマイクロ チャネル内のバイオ フィルムを育成のユニークなアプローチに焦点を当てるTOF-SIMS 解析。バイオ フィルムが以前のみ真空ベース表面分析技術のチャンバ内でサンプルを乾燥する限定されていました、ライブ バイオ フィルムの詳細な EPS とバイオ化学マッピング今入手できますその場でこの新しい機能であるためです。

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	<tbody>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">ToF-SIMS</td>
			<td style="width:260px;">IONTOF</td>
			<td style="width:123px;">TOF.SIMS 5</td>
			<td style="width:1675px;">Resolution:&#62;10,000 m/&#916;m for mass resolution;&#62;4,000 m/&#916;m for high spatial resolution</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">System for Analysis at the Liquid Vacuum Interface (SALVI)</td>
			<td style="width:260px;">Pacific Northwest National Laboratory</td>
			<td style="width:123px;">N/A</td>
			<td>SALVI is a unique, self-contained, portable analytical tool that, for the first time, enables vacuum based scientific instruments such as time-of-flight secondary ion mass spectrometry (ToF-SIMS) to analyze liquid surfaces in their natural state at the molecular level.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">-80&#176;C Freezer</td>
			<td style="width:260px;">New Brunswick Scientific</td>
			<td style="width:123px;">N/A</td>
			<td>U410 Premium Energy Efficient Ultra-Low Temperature Freezer</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">4&#176;C Refrigerator</td>
			<td style="width:260px;">BioCold Scientific</td>
			<td style="width:123px;">N/A</td>
			<td>COLDBOX1</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Orbital Shaker</td>
			<td style="width:260px;">New Brunswick Scientific</td>
			<td style="width:123px;">N/A</td>
			<td style="width:1675px;">Innova 4900 Multi-Tier Environmental Shaker, set at 30 degrees Celsius for serum bottle and flask culturing, set at 150rpm.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Syringe Pump</td>
			<td style="width:260px;">Cole-Parmer</td>
			<td style="width:123px;">EW-74905-02</td>
			<td>Cole-Parmer Syringe Pump, Infusion Only, Touchscreen Control 74905-02, used for injecting liquid into the tubing system and SALVI at a constant flowrate.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Incubator</td>
			<td style="width:260px;">Barnstead International</td>
			<td style="width:123px;">LT1465X3</td>
			<td>Lab-Line incubator, set at 30 degrees Celsius for plate culturing.</td>
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			<td height="20" style="width:389px;height:20px;">Autoclave</td>
			<td style="width:260px;">Getinge</td>
			<td style="width:123px;">533LS</td>
			<td>Used to sterilize PEEK fittings, tubing systems, serum vials, and medium. Model 533LS Vacuum Steam Sterilizer</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Spectrophotometer</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">4001-000</td>
			<td>GENESYS 20 spectrophotometer for OD600 readings of cuvettes for growth curves.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Biological Safety Cabinet</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">1385</td>
			<td>1300 Series AZ Biological Safety Cabinet</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Fluorescence Microscope</td>
			<td style="width:260px;">Nikon</td>
			<td style="width:123px;">N/A</td>
			<td>Nikon OPTIPHOT-2 fluorescence microscope with camera and super high pressure mercury lamp power supply.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">pH Meter</td>
			<td style="width:260px;">Mettler Toledo</td>
			<td style="width:123px;">51302803</td>
			<td>Used to test the pH of the &#8220;nanowires&#8221; medium after finished and before autoclaving.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">PEEK Union</td>
			<td style="width:260px;">Valco</td>
			<td style="width:123px;">ZU1TPK</td>
			<td>For connecting the inlet and outlet of SALVI, the syringe to the tubing system, and the inlet of the SALVI to the drip chamber of the tubing system.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">5 Axes Sample Stage</td>
			<td style="width:260px;">IONTOF</td>
			<td style="width:123px;">N/A</td>
			<td>Stage is self-made for mounting SALVI in ToF-SIMS.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Barnstead Nanopure Water Purification System</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">D11921</td>
			<td>ROpure LP Reverse Osmosis filtration module (D2716)</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Pipette</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">21-377-821</td>
			<td>Range: 100 to 1,000 &#181;L.</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Pipette Tip</td>
			<td style="width:260px;">Neptune</td>
			<td style="width:123px;">2112.96.BS</td>
			<td>1,000 &#181;L pipette tips</td>
		</tr>
		<tr height="20">
			<td height="20" style="width:389px;height:20px;">Razor Blade Handle</td>
			<td style="width:260px;">Stanley</td>
			<td style="width:123px;">N/A</td>
			<td>Stanley Bostitch Razor Blade Scraper with 5 Single-Edge Blades, used for cutting PTFE tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Syringe</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">309659</td>
			<td>1 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Syringe</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">309657</td>
			<td>3 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Syringe</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">309646</td>
			<td>5 mL; Used for making the drip chamber</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Syringe</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">309604</td>
			<td>10 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Syringe</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">302830</td>
			<td>20 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Disposable Pipette</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">13-678-11</td>
			<td>25 mL Fisherbrand&#8482; Sterile Polystyrene Disposable Serological Pipets with Magnifier Stripe, for filling serum bottles.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Electric Pipette Filler</td>
			<td style="width:260px;">Pipet-aid</td>
			<td style="width:123px;">P-57260</td>
			<td>Vacuum pressure electric serological pipette filler</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Serum Bottle</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">33109-U</td>
			<td>Holds approximately 69 mL of liquid for culture growth, optimum for use of 20mL culture per bottle.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Anaerobic Culture Tube</td>
			<td style="width:260px;">VWR</td>
			<td style="width:123px;">89167-178</td>
			<td>Anaerobic Tubes, 18 x 150 mm, Supplied with 20 mm Blue Butyl Rubber Stopper and Aluminum Seal.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Rubber Stopper</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">27235-U</td>
			<td>Silicone stopper, used for sealing serum bottles and for creating the tubing system/drip chamber.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Aluminum Crimp Seal (without septum)</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">27227-U</td>
			<td>Aluminum seal for top of serum bottle for use with serum bottle crimper.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Serum Bottle Aluminum Seal Crimper</td>
			<td style="width:260px;">Wheaton</td>
			<td style="width:123px;">224307</td>
			<td>30 mm crimper with standard seal.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">PTFE Tubing</td>
			<td style="width:260px;">Supelco</td>
			<td style="width:123px;">58697-U</td>
			<td>1.58 mm OD x 0.5 mm ID 50 ft. PTFE Teflon tubing, used for creating the tubing system.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Disposable Cuvettes</td>
			<td style="width:260px;">GMBH</td>
			<td style="width:123px;">759085D</td>
			<td>1.5 Ml for use with spectrophotometer.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Needle</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">303015</td>
			<td>22G; used for serum bottle injection.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Needle</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">305120</td>
			<td>23G; used for punching-through rubber stopper to create drip tubing system.</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Shewanella oneidensis MR-1 with GFP</td>
			<td style="width:260px;">N/A</td>
			<td style="width:123px;">N/A</td>
			<td>Matthysse AG, Stretton S, Dandie C, McClure NC, &#38; Goodman AE (1996) Construction of GFP vectors for use in Gram-negative bacteria other than Escherichia coli. FEMS Microbiol Lett 145(1):87-94.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Ethanol</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">&#160;S25310A</td>
			<td>95% Denatured</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">TSA</td>
			<td style="width:260px;">BD</td>
			<td style="width:123px;">212305</td>
			<td>Tryptic soy agar for culturing the model organism (S. oneidensis) used in this protocol</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">PIPES Buffer</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">P-1851</td>
			<td>Used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Hydroxide</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">S-5881</td>
			<td>Used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Ammonium Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">A-5666</td>
			<td>Used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Potassium Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">P-4504</td>
			<td>Used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Phosphate Monobasic</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">S-9638</td>
			<td>Used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Chloride</td>
			<td style="width:260px;">Thermo Fisher Scientific</td>
			<td style="width:123px;">S271-3</td>
			<td>Used for &#8220;nanowires&#8221; medium, and used to make mineral solution used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium lactate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">L-1375</td>
			<td>60%(w/w) syrup @ 98% pure, d=1.3 g/mL, 7M, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Bicarbonate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">S-5761</td>
			<td>Used to make ferric NTA solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Nitrilotriacetic Acid Trisodium Salt</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">N-0253</td>
			<td>Used to make ferric NTA solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Iron (III) Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">451649</td>
			<td>Used to make ferric NTA solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Magnesium Sulfate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">208094</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Manganese (II) Sulfate Monohydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">M-7634</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Iron(II) Sulfate Heptahydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">215422</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Calcium Chloride Dihydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">223506</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Cobalt(II) Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">60818</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Zinc Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">229997</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Copper(II) Sulfate Pentahydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">C-8027</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Aluminum Potassium Sulfate Dodecahydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">237086</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Boric Acid</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">B-6768</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Molybdate Dihydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">331058</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Nickel(II) Chloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">339350</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Sodium Tungstate Dihydrate</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">14304</td>
			<td>Used to make minerals solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">D-Biotin</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">47868</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Folic Acid</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">F-7876</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Pyridoxine Hydrochloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">P-9755</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Riboflavin (B2)</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">47861</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Thiamine Hydrochloride</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">T-4625</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Nicotinic Acid</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">N4126</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">D-Pantothenic Acid Hemicalcium Salt</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">21210</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Vitamin B12</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">V-2876</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">4-Aminobenzoic Acid</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">A-9878</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:389px;height:21px;">Thioctic Acid</td>
			<td style="width:260px;">Sigma</td>
			<td style="width:123px;">T-1395</td>
			<td>Used to make vitamin solution, used for &#8220;nanowires&#8221; medium {Hill, E.A. 2007}</td>
		</tr>
	</tbody>
</table>

in situ characterization of shewanella oneidensis mr1 biofilms by salvi and tof-sims

細菌バイオ フィルムの自主制作に細胞外高分子物質 (EPS) と環境微生物学での自分の役割を理解する研究が大幅に表面関連のコミュニティであります。本研究は、バイオ フィルムの添付ファイル分析液体真空インターフェイス (サルヴィ) でのシステムを開拓し、飛行時間二次イオン質量分析 (TOF-SIMS) によってその場で生きているバイオ フィルムの化学マッピングを実現する方法をについて説明します。これは外側と専門のセットアップでは、サルヴィ チャネル内の両方の細菌の培養と同様、バイオ フィルムの存在と TOF-SIMS 解析の前に厚さを検出する光イメージング技術です。我々 の結果は自然な水和状態でシェワネラバイオ フィルムの特徴的なピークを表示によってローカライズされた水クラスター環境、EPS の強調表示のフラグメントは、同じバイオ フィルムから大幅に異なって脱水状態です。これらの結果は、真空ベースの化学イメージング装置とその場観察バイオ イメージングは、サルヴィの画期的な機能を示しています。

これらの代表的な結果は、TOF-SIMS を通じて得られた添付のバイオ フィルムの化学物質のプロファイルを識別して、解釈方法を示すのに役立ちます。質量スペクトル TOF-SIMS データ集録、簡潔に手順セクションで強調表示を印刷後各それぞれ m/z 値識別情報を割り当てるためにピークの同定を実施しなければなりません。これは、細菌や様々 な水のクラスター、脂肪酸?...

Watch this Scientific Journal Video about In Situ Characterization of Shewanella oneidensis MR1 Biofilms by SALVI and ToF-SIMS at JoVE.com

In Situ Characterization of Shewanella oneidensis MR1 Biofilms by SALVI and ToF-SIMS

マイクロ流体リアクター、液体真空界面解析システムにより水和状態の化学マッピングのための in situ飛行時間二次イオン質量分析のためのバイオ フィルムの成長のための手法を提案します。緑色蛍光タンパク質を持つシェワネラ oneidensis氏 1 は、モデルとして使用されました。

その場でサルヴィ、TOF-SIMSシェワネラ oneidensis MR1 バイオ フィルムの特性

Bacterial biofilms are surface-associated communities that are vastly studied to understand their self-produced extracellular polymeric substances (EPS) ...

The overall goal of this procedure is to culture bacteria in the system for analysis at the liquid vacuum interface, or SALVI, for subsequent imagining of the biofilms in their natural hydrated state with vacuum-based techniques such as Time-of-Flight Secondary Ion Mass Spectrometry. We present a method that helps answer key questions in environmental microbiology by visualizing the chemical-spatial distribution of living biofilms using imaging mass spectrometry like SIMS. The main advantage of this technique is that we can introduce biofilms alive to a mass spectrometer and many other analytical platforms to obtain multiplex measurements.Although Shewanella oneidensis is used in this video, this method can also be applied to anaerobic chambers for biofilm growth. The latter requires a more stringent growth environment. Visual demonstration of this method is critical as the tubing setup steps are difficult to learn.Specific placement of tubing and care are necessary when transferring to the biosafety cabinet. Using a 22 gauge needle, carefully punch a hole through a serum bottle rubber stopper. Cut 20 inches of 1/32 inch PTFE tubing with a razor, such that one end of the tubing is pointed.Using the pointed end, thread the PTFE tubing through the hole in the top portion of the rubber stopper. Push the tube through until roughly two centimeters is through the stopper and cut the pointed end of the PTFE tube with a razor to have a flat end. Remove the plunger of a five milliliter syringe and fit the rubber stopper into the open end of the syringe.The two centimeter end of PTFE tubing is within the barrel of the syringe. This is the bubble trap. Wrap the bubble trap with aluminum foil and secure it with autoclave tape.Then autoclave the foil package to sterilize the tubing with a gravity cycle at 121 degrees celsius for 30 minutes. Store the sealed foil package at room temperature until it is ready for use. This procedure specifically covers culturing a Shewanella oneidensis MR-1 biofilm at room temperature after 24 hours of incubation.However, different strains of bacteria require different growth conditions and lengths to achieve a desirable thickness for analysis. The SALVI tubing system is made up of the syringe containing the liquid reservoir, which is attached with a PEEK injector to the PTFE tubing. This is attached to the drip chamber, which has a PEEK injector fitting attached to the SALVI inlet tubing.The drip chamber is also attached to the outlet container that the SALVI outlet tubing is sealed to. To promote sterility of the tubing system, detach the syringe from the syringe pump by unscrewing the metal syringe holder. Then bring the syringe with the attached tubing to a sterile biological safety cabinet.Use a new SALVI device and attach a PEEk fitting and injector to one end of a PTFE tubing. SALVI devices are prepared fresh for each experiment following the device fabrication, which is detailed in previous research and patents. Next, aspirate two milliliters of 70%ethanol and deionized water into a syringe and connect the syringe to the PEEK fitting on the SALVI.After connecting the syringe to a syringe pump and attaching the outlet of the SALVI to a waste bottle, allow the ethanol solution to run through the SALVI at 20 microliters per minute for one and a half hours. Then aspirate four milliliters of sterilized deionized water into a syringe and connect it to the inlet of the same SALVI. After connecting the syringe to a syringe pump, allow the water to run through the SALVI at 20 microliters per minute for at least three hours.Within a biosafety cabinet, open the prepared foil packet and connect a sterile PEEK injector and fittings to the end of the PTFE tubing. Aspirate approximately three milliliters of sterile medium into a syringe and attach it to the PTFE tube. Invert the five milliliter drip chamber and using a sterile syringe inject the growth medium into the drip chamber until it reaches a total volume of one milliliter.Next, connect the end of the five milliliter syringe drip chamber to the inlet of the SALVI. Aspirate 10 milliliters of growth medium into a sterile syringe and connect it to the inlet of the drip tubing. Allow the drip tubing to be vertical.The syringe pump should be placed on an elevated surface with drip tubing secured by tape and with the SALVI secured on a flat surface below. Cover the outlet bottle with foil or plastic paraffin film to minimize the probability of medium contamination. Allow the medium to run through the SAVLI at 20 microliters per minute for 12 hours.This ensures that all traces of ethanol have been removed from the microchannel and tubing of the SALVI before inoculating with bacteria. For added protection, keep the SALVI microchannel chamber within a sterilized petri dish with the sides cut to fit the inlet and outlet tubing. Additionally, keep tape always on the window to protect the silicone nitride membrane and channel prior to image analysis.We move the whole tubing system from the syringe pump to bring it to a sterilized biosafety cabinet. Additionally, remove the serum bottle from the incubator that has been inoculated as described in the text protocol. Clean the surface of the serum bottle stopper with 70%ethanol to prevent any contamination.Then use a sterile syringe with an attached sterile 22 gauge needle to extract four milliliters of bacteria from the bottle. Detach the SALVI from the five milliliter chamber of the drip tubing and connect the syringe with inoculated medium directly to the inlet of the SALVI. Leave the drip tubing within the sterile aluminum foil packet or within the biosafety cabinet.Attach the syringe with the SALVI and outlet bottle to the syringe pump. Run at 20 microliters per minute for three hours to inoculate the channel of the SALVI and allow for a multiple volume changes of the liquid contained within the SALVI. Following inoculation, disconnect the syringe with the SALVI from the syringe pump and bring it to the biosafety cabinet.Attach the inlet of the SALVI back to the five milliliter drip chamber. Then aspirate 20 milliliters of growth medium into a sterile syringe and attach it to the inlet of the drip tubing. The flow rate should never be increased or decreased, as changing the flow rate would create sheer stress within the channel and detach the biofilm.To avoid flow rate changes ensure there are no bubbles in the tubing the day before SIMS. It is important to be cautious of bubbles within the bottom of the five milliliter drip tubing where it connects to the SALVI tubing. Fresh medium can be injected into the end of the PEEK injector to ensure that no air will be forced into the SALVI.Bring the tubing attached to the SALVI from the biosafety cabinet to the syringe pump. Allow the medium to pass through the tubing at at rate of two microliters per minute for six to ten days. Refill the SALVI with fresh medium as it runs out within the biosafety cabinet by attaching a new sterile syringe filled with 10 milliliters of growth medium to the SALVI.Using light microscopy or confocal fluorescence imaging, the presence of a biofilm can be detected on the window of the SALVI. Shown here is the negative mass spectrum of a hydrated Shewanella oneidensis MR-1 biofilm within the microfluidic channel. It's obtained by NC2 liquid Time-of-Flight Secondary Ion Mass Spectrometry.The deionized water spectrum is also shown. A comparison of 2-D images of peaks of interest in the MR-1 biofilm to deionized water are shown. Once interesting m over z values can be identified from studying the mass spectra potential peak identification is performed through the software library, in lab reference spectra, and a literature survey.Once mastered, this technique takes three days to set up, in addition to the specific time required for the biofilm to grow within the microchannel, which is about five to six days at room temperature for Shewanella. After watching this video, you should have a good understanding of how to grow insatiable biofilms in SALVI devices. Generally, individuals new to this method will struggle because bubbles may form within the growth medium due to bacteria growth.It is necessary to get rid of bubbles before the SIMS analysis. While attempting this procedure, it's important to remember to never change the flow rate after inoculation of bacteria is completed. Even in fluorofil dying steps before optical imaging.Following this procedure, other methods like confocal fluorescence imaging can be performed in order to answer additional questions, like how thick the biofilm was at the time the SIMS took place. After it's development, the technique paved the way for researchers in analytical chemistry and microbiology to explore the chemical profiles of biofilms in their naturally hydrated states.

Research

JoVE Journal

Chemistry

In Situ Characterization of Shewanella oneidensis MR1 Biofilms by SALVI and ToF-SIMS

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

その場 SIMSおよび質量選択されたイオンのソフトランディングで調製し明確に定義された表面の赤外分光法における

Soft landing of mass-selected ions onto surfaces is a powerful approach for the highly-controlled preparation of materials that are inaccessible using ...

化学

Assessment of Boron Doped Diamond Electrode Quality and Application to In Situ Modification of Local pH by Water Electrolysis

Assessment of Boron Doped Diamond Electrode Quality and Application to In Situ Modification of Local pH by Water Electrolysis

ホウ素ドープダイヤモンド電極品質とアプリケーションの評価<em&gt;その場で</em&gt;水電解による局所のpHの変更

Boron doped diamond (BDD) electrodes have shown considerable promise as an electrode material where many of their reported properties such as extended ...

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

高速液体クロマトグラフィー結合フローシンチレーションにより放射性標識およびホスホイノシチドの細胞レベルの定量化

Phosphoinositides (PtdInsPs) are essential signaling lipids responsible for recruiting specific effectors and conferring organelles with molecular ...

In Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS

In Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS

サルヴィとTOF-SIMSによる水中で水和タンパク質のその場キャラクタリゼーションで

This work demonstrates in situ characterization of protein biomolecules in the aqueous solution using the System for Analysis at the Liquid Vacuum ...

Characterizing Electron Transport through Living Biofilms

生きているバイオ フィルムにおける電子輸送特性評価

Here we demonstrate the method of electrochemical gating used to characterize electrical conductivity of electrode-grown microbial biofilms under ...

In Situ Characterization of Boehmite Particles in Water Using Liquid SEM

In Situ Characterization of Boehmite Particles in Water Using Liquid SEM

その場で液体の SEM を用いた水中のベーマイト粒子のキャラクタリゼーション

In situ imaging and elemental analysis of boehmite (AlOOH) particles in water is realized using the System for Analysis at the Liquid Vacuum Interface ...

DNA-magnetic Particle Binding Analysis by Dynamic and Electrophoretic Light Scattering

バインドの動的・電気泳動光散乱による解析 DNA 磁性粒子

Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the ...

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

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

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 ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

原金ナノ粒子合成による急速なナノプローブ信号強調

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...

Photoelectron Imaging of Anions Illustrated by 310 Nm Detachment of F&#8722;

F− 310 Nm 剥離によって示されている陰イオンの光電子イメージング

Anion photoelectron imaging is a very efficient method for the study of energy states of bound negative ions, neutral species and interactions of unbound ...

その場でサルヴィ、TOF-SIMSシェワネラ oneidensis MR1 バイオ フィルムの特性

その場でサルヴィ、TOF-SIMSシェワネラ oneidensis MR1 バイオフィルムの特性