Name: investigating the detrimental effects of low pressure plasma sterilization on the survival of bacillus subtilis spores using live cell microscopy
Uploaded: 2017-11-30T14:00:00
Description: Watch this Scientific Journal Video about Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy at JoVE.com

作者感谢安德莉亚在这项工作的部分和 Nikea 在视频拍摄期间对她的帮助的出色的技术援助。我们还要感谢莱尔对她慷慨捐赠的枯草芽孢杆菌菌株: LAS72 和 LAS24。这项工作得到了部分支持, 从德国研究基金会 (DFG) Paketantrag (PlasmaDecon 柏 728) 到 PA (AW 7/3-1) 和 RM (MO 2023/2-1) 和 dlr 赠款 dlr-FuW-Projekt ISS 生活, 射频 FuW, Teilprogramm 475 (f.m F, 大副和 rm)。F.M.F. 由亥姆霍兹空间生命科学研究学校 (SpaceLife) 的博士奖学金在德国的科隆, 德国航空航天中心 (DLR) 支持, 由亥姆霍兹协会 (亥姆霍兹-礼俗) 提供经费在六年期间 (格兰特 No。VH-KO-300) 并获得了 DLR 的额外资金, 包括航空航天执行委员会和航空航天医学研究所。这项研究的结果将被包括在博士论文的费利克斯 m。

1.枯草芽孢杆菌孢子的生产和纯化
<ol><li>为孢子生产, 转移各自的B. 枯草芽孢杆菌菌株的5毫升隔夜文化, 补充适当的抗生素, 到200毫升双强度液体谢弗产培养基 (每公升 16 g 营养素肉汤, 氯化钾 2 g, 0.5 g MgSO4* 7 H2O, 2 mL 1 M Ca (不3)2, 2 毫升 0.1 M MnCl2 * 4 h2O, 2 ml 1 mM FeSO4, 2 ml 50% (w/v) 葡萄糖18), 并培养它与蓬勃发展的?...

<ol>
	<li>Nicholson, W. L., Schuerger, A. C., Race, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migrating+microbes+and+planetary+protection.">Migrating microbes and planetary protection.</a> Trends Microbiol. 17, 389-392 (2009).</li><li>COSPAR. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=COSPAR+Planetery+Protection+Policy.">COSPAR Planetery Protection Policy.</a> Space Research Today, COSPAR's Information Bulletin. 193, 1-14 (2015).</li><li>De Geyter, N., Morent, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonthermal+plasma+sterilization+of+living+and+nonliving+surfaces.">Nonthermal plasma sterilization of living and nonliving surfaces.</a> Annu Rev Biomed Eng. 14, 255-274 (2012).</li><li>Shimizu, S., 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=Cold+atmospheric+plasma+-+A+new+technology+for+spacecraft+component+decontamination.">Cold atmospheric plasma - A new technology for spacecraft component decontamination.</a> Planet. Space Sci. 90, 60-71 (2014).</li><li>Lerouge, S., Fozza, A. C., Wertheimer, M. R., Marchand, R., Yahia, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sterilization+by+Low-Pressure+Plasma:+The+Role+of+Vacuum-Ultraviolet+Radiation.">Sterilization by Low-Pressure Plasma: The Role of Vacuum-Ultraviolet Radiation.</a> Plasma Polym. 5, 31-46 (2000).</li><li>Rossi, F., Kyli&#225;n, O., Rauscher, H., Gilliland, D., Sirghi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+low-pressure+plasma+discharge+for+the+decontamination+and+sterilization+of+medical+devices.">Use of a low-pressure plasma discharge for the decontamination and sterilization of medical devices.</a> Pure Appl. Chem. 80, 1939-1951 (2008).</li><li>Halfmann, H., Hauser, J., Awakowicz, P., Koller, M., Esenwein, S. 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+double+inductively+coupled+low-pressure+plasma+for+sterilization+of+medical+implant+materials.">A double inductively coupled low-pressure plasma for sterilization of medical implant materials.</a> Biomed Tech (Berl). 53, 199-203 (2008).</li><li>Halfmann, H., Denis, B., Bibinov, N., Wunderlich, J., Awakowicz, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+the+most+efficient+VUV/UV+radiation+for+plasma+based+inactivation+of+Bacillus+atrophaeus+spores.">Identification of the most efficient VUV/UV radiation for plasma based inactivation of Bacillus atrophaeus spores.</a> J. Phys. D: Appl. Phys. 40, 5907 (2007).</li><li>Vaishampayan, P., 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=Bacillus+horneckiae+sp.+nov.,+isolated+from+a+spacecraft-assembly+clean+room.">Bacillus horneckiae sp. nov., isolated from a spacecraft-assembly clean room.</a> Int J Syst Evol Microbiol. 60, 1031-1037 (2010).</li><li>Mandic-Mulec, I., Stefanic, P., van Elsas, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecology+of+Bacillaceae.">Ecology of Bacillaceae.</a> Microbiol Spectr. 3, (2015).</li><li>Alekhova, T. 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=Diversity+of+bacteria+of+the+genus+Bacillus+on+board+of+international+space+station.">Diversity of bacteria of the genus Bacillus on board of international space station.</a> Dokl Biochem Biophys. 465, 347-350 (2015).</li><li>Claus, D., Bekerley, R. C. W., Sneath, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genus+Bacillus+Cohn+1872.">Genus Bacillus Cohn 1872.</a> Bergey's manual of systematic bacteriology. 2, 1105-1141 (1986).</li><li>Setlow, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spore+Resistance+Properties.">Spore Resistance Properties.</a> Microbiol Spectr. 2, (2014).</li><li>Setlow, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spores+of+Bacillus+subtilis:+their+resistance+to+and+killing+by+radiation,+heat+and+chemicals.">Spores of Bacillus subtilis: their resistance to and killing by radiation, heat and chemicals.</a> J Appl Microbiol. 101, 514-525 (2006).</li><li>Roth, S., Feichtinger, J., Hertel, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+Bacillus+subtilis+spore+inactivation+in+low-pressure,+low-temperature+gas+plasma+sterilization+processes.">Characterization of Bacillus subtilis spore inactivation in low-pressure, low-temperature gas plasma sterilization processes.</a> J Appl Microbiol. 108, 521-531 (2010).</li><li>Roth, S., Feichtinger, J., Hertel, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Response+of+Deinococcus+radiodurans+to+low-pressure+low-temperature+plasma+sterilization+processes.">Response of Deinococcus radiodurans to low-pressure low-temperature plasma sterilization processes.</a> J Appl Microbiol. 109, 1521-1530 (2010).</li><li>Setlow, B., Setlow, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+DNA+repair+in+Bacillus+subtilis+spore+resistance.">Role of DNA repair in Bacillus subtilis spore resistance.</a> J Bacteriol. 178, 3486-3495 (1996).</li><li>Schaeffer, P., Millet, J., Aubert, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catabolic+repression+of+bacterial+sporulation.">Catabolic repression of bacterial sporulation.</a> Proc. Natl. Acad. Sci. 54, 704-711 (1965).</li><li>Simmons, L. 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=Comparison+of+responses+to+double-strand+breaks+between+Escherichia+coli+and+Bacillus+subtilis+reveals+different+requirements+for+SOS+induction.">Comparison of responses to double-strand breaks between Escherichia coli and Bacillus subtilis reveals different requirements for SOS induction.</a> J Bacteriol. 191, 1152-1161 (2009).</li><li>Raguse, 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=Improvement+of+Biological+Indicators+by+Uniformly+Distributing+Bacillus+subtilis+Spores+in+Monolayers+To+Evaluate+Enhanced+Spore+Decontamination+Technologies.">Improvement of Biological Indicators by Uniformly Distributing Bacillus subtilis Spores in Monolayers To Evaluate Enhanced Spore Decontamination Technologies.</a> Appl Environ Microbiol. 82, 2031-2038 (2016).</li><li>Horneck, G., 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=Protection+of+bacterial+spores+in+space,+a+contribution+to+the+discussion+on+Panspermia.">Protection of bacterial spores in space, a contribution to the discussion on Panspermia.</a> Orig Life Evol Biosph. 31, 527-547 (2001).</li><li>Opretzka, J., Benedikt, J., Awakowicz, P., Wunderlich, J., Keudell, A. v. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+chemical+sputtering+during+plasma+sterilization+of+Bacillus+atrophaeus.">The role of chemical sputtering during plasma sterilization of Bacillus atrophaeus.</a> J. Phys. D: Appl. Phys. 40, 2826 (2007).</li><li>Stapelmann, 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=Utilization+of+low-pressure+plasma+to+inactivate+bacterial+spores+on+stainless+steel+screws.">Utilization of low-pressure plasma to inactivate bacterial spores on stainless steel screws.</a> Int. J. Astrobiol. 13, 597-606 (2013).</li><li>Raguse, 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=Understanding+of+the+importance+of+the+spore+coat+structure+and+pigmentation+in+the+Bacillus+subtilis+spore+resistance+to+low-pressure+plasma+sterilization.">Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low-pressure plasma sterilization.</a> J. Phys. D: Appl. Phys. 49, 285401 (2016).</li><li>Pandey, R., 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=Live+cell+imaging+of+germination+and+outgrowth+of+individual+Bacillus+subtilis+spores;+the+effect+of+heat+stress+quantitatively+analyzed+with+SporeTracker.">Live cell imaging of germination and outgrowth of individual Bacillus subtilis spores; the effect of heat stress quantitatively analyzed with SporeTracker.</a> PloS one. 8, e58972 (2013).</li></ol>

使用低温、低压等离子体进行表面灭菌是一种很有前途的替代品, 例如用电离辐射、化学药品 (例如,2O2之类的气体进行处理) 和传统的灭菌程序环氧乙烷) 或干燥和潮湿的热量23。一般的杀菌方法主要是提供有效的灭菌, 但已知会影响处理过的材料, 并对操作者构成潜在的风险。低压等离子体提供了一个快速和均匀的生物失活, 使用的成分, 如紫外线光子, 自由...

太空探索的一个主要目标是在太阳系的其他行星体和卫星上寻找生命形式和生物分子的特征。将地球起源的微生物或生物分子转移到重要的勘探领域, 对火星和欧罗巴1等行星机构的生命探测任务的发展和完整性产生了特别的影响。1967年由空间研究委员会设立的《行星保护国际准则》对载人和机器人任务向其他行星、卫星、小行星和其他天体实施了严格的管制, 并对在发射前对航天器和关键硬件部件进行清洗和灭菌, 以消除对陆地微生物的污染, 防止天体的交叉污染2。在过去的十年中, 非热等离子体的应用在生物医学和营养学研究以及航天应用中得到了广泛的关注,3,4,5。等离子体灭菌是一种有前途的替代传统的杀菌方法, 因为它提供了快速和有效的微生物失活6, 同时对敏感和热不稳定材料温和。等离子体放电含有活性剂的混合物, 如自由基、带电粒子、中性/激发原子、紫外线 (UV) 中的光子和真空紫外 (真空) 谱, 导致快速的微生物失活3。本研究采用双电感耦合低压等离子体产生的低压等离子体 (DICP) 源7,8灭活枯草芽孢杆菌孢子在玻璃测试表面上分布。
家庭的革兰氏阳性菌Bacillaceae广泛分布在土壤、沉积物和空气的自然栖息地以及在不寻常的环境中, 如洁净室设施和国际空间站9,10 ,11。最明显的特点,芽孢杆菌是能力形成高抗性休眠孢子 (以下简称孢子) 生存不利的条件, 如养分枯竭12。孢子通常比它们的营养细胞相对于各种处理和环境压力更具抵抗力, 包括热、紫外线、γ辐照、脱水、机械破坏和有毒化学物质, 如强氧化剂或pH 变化的试剂 (在参考文献13,14), 因此是测试微生物失活方法效率的理想对象。由于基因组 DNA 是细菌的等离子体治疗的主要目标15,16, 修复的血浆诱导的 dna 损伤 (如 dna 双链断裂) 孢子复活是至关重要的生存细菌13, 17。
因此, 我们研究了孢子萌发能力和 dna 修复在孢子萌发和生长过程中的作用, 处理后的孢子与低压力氩等离子体的个别孢子和它们的表达荧光标记 dna 修复蛋白质 RecA 与时间分辨共焦荧光显微镜。我们给出了在单分子膜中制备枯草芽孢杆菌孢子的步骤说明, 以实现重现性试验结果, 用低压等离子体处理芽孢膜灭菌, 制备等离子体处理的孢子超微结构评价使用扫描电子显微镜 (SEM) 和现场细胞显微镜分析的个人孢子的水平与监测的活性 DNA 修复过程中发生的细胞内反应等离子体治疗。

<table>
	<tbody>
		<tr>
			<td>Two substance nozzle (model 970-8)</td>
			<td>Schlick</td>
			<td>14,404</td>
			<td>230 V, 50 Hz, D 4.484/8, 0.8 mm bore diameter</td>
		</tr>
		<tr>
			<td>Luria Bertani Medium</td>
			<td>Sigma Aldrich</td>
			<td>70122-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Tube connectors</td>
			<td>Festo</td>
			<td>n/a</td>
			<td>G 1/8</td>
		</tr>
		<tr>
			<td>Magnetvalve DO35-3/2NC-G018-230AC</td>
			<td>Bosch Rexroth</td>
			<td>820005100</td>
			<td></td>
		</tr>
		<tr>
			<td>PLN Polyamid tube</td>
			<td>Festo</td>
			<td>558206</td>
			<td>d = 6 mm</td>
		</tr>
		<tr>
			<td>Glass slides</td>
			<td>VWR</td>
			<td>48300-026</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric Timer 550-2-C</td>
			<td>Gefran</td>
			<td>F000074</td>
			<td>220 V</td>
		</tr>
		<tr>
			<td>attofluor cell chamber</td>
			<td>Menzel, Fisher Ref.</td>
			<td>3406816</td>
			<td>d=25 mm, round</td>
		</tr>
		<tr>
			<td>MgSO4*7 H2O</td>
			<td>Sigma Aldrich</td>
			<td>13152</td>
			<td></td>
		</tr>
		<tr>
			<td>Ca(NO3)2</td>
			<td>Sigma Aldrich</td>
			<td>202967</td>
			<td></td>
		</tr>
		<tr>
			<td>MnCl2 * 4 H2O</td>
			<td>Sigma Aldrich</td>
			<td>244589</td>
			<td></td>
		</tr>
		<tr>
			<td>FeSO4 * 7H2O</td>
			<td>AppliChem</td>
			<td>13446-34-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Merck</td>
			<td>215422</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma Aldrich</td>
			<td>P9541-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Nutrient Broth (NB)</td>
			<td>Merck</td>
			<td>105443</td>
			<td></td>
		</tr>
		<tr>
			<td>Luria-Bertani (LB)</td>
			<td>Merck</td>
			<td>110283</td>
			<td></td>
		</tr>
		<tr>
			<td>96-wellplate</td>
			<td>ThermoFisher</td>
			<td>243656</td>
			<td></td>
		</tr>
		<tr>
			<td>Zeiss LSM 780, Axio Observer Z1</td>
			<td>Carl Zeiss Microscopy GmbH</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Leo 1530 Gemini</td>
			<td>Carl Zeiss Microscopy GmbH</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>ZEN 2 and ZEN lite 2012 (Software)</td>
			<td>Carl Zeiss Microscopy GmbH</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>SigmaPlot, version 13.0 (Statistic software)</td>
			<td>Systat GmbH, Erkrath, Germany</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Attofluor cell chamber</td>
			<td>Invitrogen</td>
			<td>A7816</td>
			<td></td>
		</tr>
		<tr>
			<td>&#181;-Dish 35 mm, high Grid-500 Glass Bottom</td>
			<td>ibidi</td>
			<td>81168</td>
			<td></td>
		</tr>
	</tbody>
</table>

investigating the detrimental effects of low pressure plasma sterilization on the survival of bacillus subtilis spores using live cell microscopy

等离子体灭菌是一种有前途的替代传统的杀菌方法的工业, 临床和航天的目的。低压等离子体 (LPP) 放电含有广泛的活性物种, 导致快速的微生物失活。为了研究 LPP 杀菌的效率和机制, 我们使用试验生物体的孢子枯草芽孢杆菌, 因为它们对常规灭菌程序的抵抗力非常高。我们描述了枯草芽孢杆菌的生产孢子单分子膜, 低压等离子体在双电感耦合等离子体反应器中的杀菌过程, 用扫描电子显微镜 (SEM) 表征孢子形态, 以及用活细胞显微镜分析孢子的萌发和生长。血浆物种的一个主要目标是基因组材料 (dna) 和修复的等离子体诱导 DNA 损伤孢子复活是至关重要的生存的有机体。在这里, 我们研究了孢子萌发能力和 dna 修复在孢子萌发和生长后 LPP 的作用, 通过跟踪荧光标记的 dna 修复蛋白 (RecA) 与时间分辨共聚焦荧光显微镜。经过治疗和未经处理的孢子单膜激活发芽和可视化的倒置共聚焦活细胞显微镜随时间的反应, 个别孢子。我们的观察表明, 萌发和超过孢子的分数取决于 LPP 的持续时间, 在120s 后达到最低限度. RecA-YFP (黄色荧光蛋白) 荧光检测仅在少数孢子和发展在所有超过细胞, LPP 处理孢子略有升高。此外, 从 LPP 处理的孢子中获得的一些植物性细菌在细胞质中呈上升趋势, 并趋于溶解。所描述的个体孢子分析方法可以作为研究孢子萌发和生长其他方面的典范。

等离子体处理的枯草芽孢杆菌孢子的存活
等离子体处理的枯草芽孢杆菌在本研究中使用的孢子显示, 随着等离子体处理时间的延长, 存活率下降 (图 2)。表达recA-基因的菌株的孢子 YFP 显示了类似于野生型菌株孢子的存活曲线, 表明基因修饰对细菌活力没有显著影响。与野生型菌株的孢子?...

Watch this Scientific Journal Video about Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy at JoVE.com

Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy

用活细胞显微镜研究低压等离子体灭菌对枯草芽孢杆菌孢子存活的不利影响

这项协议说明了评估在用低压等离子体处理后恢复枯草芽孢杆菌孢子后, 监测活力参数和 DNA 修复过程的相关性所需的重要连续步骤。荧光标记的 DNA 修复蛋白质通过时间分辨共焦显微镜和扫描电子显微镜。

Plasma sterilization is a promising alternative to conventional sterilization methods for industrial, clinical, and spaceflight purposes. Low pressure ...

Research

JoVE Journal

Biology

Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy

Imaging Mismatch Repair and Cellular Responses to DNA Damage in Bacillus subtilis

Imaging Mismatch Repair and Cellular Responses to DNA Damage in Bacillus subtilis

成像错配修复和DNA损伤的细胞反应枯草芽孢杆菌</em

Both prokaryotes and eukaryotes respond to DNA damage through a complex set of physiological changes. Alterations in gene expression, the redistribution ...

科研

生物学

Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1

使用内源性野生型BRCA1基因的细胞表达，确定同源重组的BRCA1基因突变的影响

The functional analysis of missense mutations can be complicated by the presence in the cell of the endogenous protein. Structure-function analyses of the ...

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

FRET显微镜在活细胞单分子生物传感器的信号事件的实时监测

F&ouml;rster resonance energy transfer (FRET) microscopy  continues to gain increasing interest as a technique for real-time monitoring  of biochemical ...

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

凋亡细胞清除过程中实时成像<em&gt;果蝇</em&gt;胚胎发育

The  proper elimination of unwanted or aberrant cells through apoptosis and  subsequent phagocytosis (apoptotic cell clearance) is crucial for normal  ...

Analyzing Craniofacial Morphogenesis in Zebrafish Using 4D Confocal Microscopy

使用4D共聚焦显微镜分析颅面形态的斑马鱼

Time-lapse imaging is a technique that allows for the direct observation of the process of morphogenesis, or the generation of shape. Due to their optical ...

Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel

植物根系的轻质板材荧光显微镜生长在一个凝胶表面

One of the key questions in understanding plant development is how single cells behave in a larger context of the tissue. Therefore, it requires the ...

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

拟南芥种子高通量灭菌的标准化方法

Arabidopsis thaliana (Arabidopsis) seedlings often need to be grown on sterile media. This requires prior seed sterilization to prevent the growth of ...

Single-cell Microfluidic Analysis of Bacillus subtilis

Single-cell Microfluidic Analysis of Bacillus subtilis

单细胞微流控芯片分析枯草芽孢杆菌

Microfluidic technology overcomes many of the limitations to traditional analytical methods in microbiology. Unlike bulk-culture methods, it offers ...

Biological Sample Preparation by High-pressure Freezing, Microwave-assisted Contrast Enhancement, and Minimal Resin Embedding for Volume Imaging

高压冷冻、微波辅助对比度增强和最小树脂嵌入在体积成像中的生物样品制备

The described sample preparation technique is designed to combine the best quality of ultrastructural preservation with the most suitable contrast for the ...

Live-Cell Imaging of the Life Cycle of Bacterial Predator Bdellovibrio bacteriovorus using Time-Lapse Fluorescence Microscopy

Live-Cell Imaging of the Life Cycle of Bacterial Predator Bdellovibrio bacteriovorus using Time-Lapse Fluorescence Microscopy

使用延时荧光显微镜对细菌捕食者 Bdellovibrio 细菌的生命周期进行活细胞成像

Bdellovibrio bacteriovorus is a small gram-negative, obligate predatory bacterium that kills other gram-negative bacteria, including harmful pathogens. ...