Name: colorimetric analysis of alkaline phosphatase activity in s. aureus biofilm
Uploaded: 2019-04-12T15:00:00
Description: Watch this Scientific Journal Video about Colorimetric Analysis of Alkaline Phosphatase Activity in S. aureus Biofilm at JoVE.com

我们感谢威廉·雷尼·哈珀学院和伊利诺伊大学提供的进行这些实验的设施。我们还感谢麦格劳·希尔基金会的慷慨支持。

1. 中等准备
<ol><li>制备1升的胰蛋白酶大豆汤 (TSB): 进入1升的蒸馏水, 加入15克的酪蛋白胰腺消化, 5 克的豆粕纸质消化, 3 克氯化钠, 2.5 克的葡萄糖, 和2.5 克的磷酸二钾。使用前消毒。</li>
	<li>对于胰蛋白酶大豆琼脂 (TSA), 在 TSB 的 1 L 中加入15克琼脂, 高压灭菌器。然后让它冷却到室温。接下来, 以每培养皿20毫升 (100 毫米 x15 毫米) 的比例倒入。</li>
	<li>生物膜培养基: 将10克葡萄糖加入蒸压 TSB 1 升。?...

<ol>
	<li>Coleman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+mechanism+of+alkaline+phosphatase.">Structure and mechanism of alkaline phosphatase.</a> Annual Review of Biophysics and Biomolecular Structure. 21, 441-483 (1992).</li><li>Sharma, U., Pal, D., Prasad, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alkaline+Phosphatase:+An+Overview.">Alkaline Phosphatase: An Overview.</a> Indian Journal of Clinical Biochemistry. 29 (3), 269-278 (2014).</li><li>Mitchell, M. 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=High+frequencies+of+elevated+alkaline+phosphatase+activity+and+rickets+exist+in+extremely+low+birth+weight+infants+despite+current+nutritional+support.">High frequencies of elevated alkaline phosphatase activity and rickets exist in extremely low birth weight infants despite current nutritional support.</a> Boston Medical Center Pediatrics. 9, 47 (2009).</li><li>Fawley, J., Gourlay, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+alkaline+phosphatase:+A+summary+of+its+role+in+clinical+disease.">Intestinal alkaline phosphatase: A summary of its role in clinical disease.</a> Journal of Surgical Research. 202 (1), 225-234 (2016).</li><li>Derman, A. I., Beckwith, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Escherichia+coli+Alkaline+Phosphatase+Localized+to+the+Cytoplasm+Slowly+Acquires+Enzymatic+Activity+in+Cells+Whose+Growth+Has+Been+Suspended:+a+Caution+for+Gene+Fusion+Studies.">Escherichia coli Alkaline Phosphatase Localized to the Cytoplasm Slowly Acquires Enzymatic Activity in Cells Whose Growth Has Been Suspended: a Caution for Gene Fusion Studies.</a> Journal of Bacteriology. 177 (13), 3764-3770 (1995).</li><li>Danikowski, K. M., Cheng, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alkaline+Phosphatase+Activity+of+Staphylococcus+aureus+grown+in+Biofilm+and+Suspension+Cultures.">Alkaline Phosphatase Activity of Staphylococcus aureus grown in Biofilm and Suspension Cultures.</a> Current Microbiology. 75, 1226-1230 (2018).</li><li>Okabayashi, K., Futai, M., Mizuno, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+Acid+and+Alkaline+Phosphatases+in+Staphylococcus+aureus.">Localization of Acid and Alkaline Phosphatases in Staphylococcus aureus.</a> Japanese Journal of Microbiology. 18 (4), 287-294 (1974).</li><li>Cheng, K. J., Costerton, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alkaline+Phosphatase+Activity+of+Rumen+Bacteria.">Alkaline Phosphatase Activity of Rumen Bacteria.</a> Applied and Environmental Microbiology. , 586-590 (1997).</li><li>Berezhetskyy, A. 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=Phosphatase+conductometric+biosensor+for+heavy-metal+ions+determination.">Phosphatase conductometric biosensor for heavy-metal ions determination.</a> Innovation and Research in Biomedical Engineering. 29 (2-3), 136-140 (2008).</li><li>Park, E. J., Kang, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+bacterial+alkaline+phosphatase+assay+for+rapid+monitoring+of+bacterial+counts+on+spinach.">The use of bacterial alkaline phosphatase assay for rapid monitoring of bacterial counts on spinach.</a> Journal of Food Science. 73 (5), M236-M238 (2008).</li><li>Barber, M., Kuper, S. W. A. <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+Staphylococcus+pyogenes+by+the+phosphatase+reaction.">Identification of Staphylococcus pyogenes by the phosphatase reaction.</a> The Journal of Pathology and Bacteriology. 63, 65-68 (1951).</li><li>Wolf, P. L., Von der Muehll, E., Ludwick, 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+new+test+to+differentiate+Serratia+from+Enterobacter.">A new test to differentiate Serratia from Enterobacter.</a> American Journal of Clinical Pathology. 56, 241-243 (1972).</li><li>Rangam, C. M., Katdare, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phosphatase+activity+of+Staphylococci+as+an+indication+of+their+pathogenicity.">Phosphatase activity of Staphylococci as an indication of their pathogenicity.</a> Indian Journal of Medical Science. 8, 610-613 (1954).</li><li>Flemming, H. -. C., 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=Biofilms:+an+emergent+form+of+bacterial+life.">Biofilms: an emergent form of bacterial life.</a> Nature Reviews Microbiology. 14, 563-575 (2016).</li><li>Hoiby, N., Bjarnsholt, T., Givskov, M., Molin, S., Ciofu, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibiotic+resistance+of+bacterial+biofilms.">Antibiotic resistance of bacterial biofilms.</a> International Journal of Antimicrobial Agents. 35, 322-332 (2010).</li><li>Ito, A., Taniuchi, A., May, T., Kawata, K., Okabe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+antibiotic+resistance+of+Escherichia+coli+in+mature+biofilms.">Increased antibiotic resistance of Escherichia coli in mature biofilms.</a> Applied and Environmental Microbiology. 75, 4093-4100 (2009).</li><li>Witherow, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Ten-Week+Biochemistry+Lab+Project+Studying+Wild-Type+and+Mutant+Bacterial+Alkaline+Phosphatase.">A Ten-Week Biochemistry Lab Project Studying Wild-Type and Mutant Bacterial Alkaline Phosphatase.</a> Biochemistry and Molecular Biology Education. 44 (6), (2016).</li><li>Henthorn, P., Zervos, P., Raducha, M., Harris, H., Kadesch, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+a+human+placental+alkaline+phosphatase+gene+in+transfected+cells:+Use+as+a+reporter+for+studies+of+gene+expression.">Expression of a human placental alkaline phosphatase gene in transfected cells: Use as a reporter for studies of gene expression.</a> Proceedings of the National Academy of Science USA. 85, 6324-6346 (1988).</li><li>Horney, B. S., Farmer, A. J., Honor, D. J., MacKenzie, A., Burton, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Agarose+gel+electrophoresis+of+alkaline+phosphatase+isoenzymes+in+the+serum+of+hyperthyroid+cats.">Agarose gel electrophoresis of alkaline phosphatase isoenzymes in the serum of hyperthyroid cats.</a> Veterinary Clinical Pathology. 23 (3), 98-102 (1994).</li><li>Farley, J. 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=Quantification+of+Skeletal+Alkaline+Phosphatase+in+Osteoporotic+Serum+by+Wheat+Germ+Agglutinin+Precipitation,+Heat+Inactivation,+and+a+Two-Site+Immunoradiometric+Assay.">Quantification of Skeletal Alkaline Phosphatase in Osteoporotic Serum by Wheat Germ Agglutinin Precipitation, Heat Inactivation, and a Two-Site Immunoradiometric Assay.</a> Clinical Chemistry. 40 (9), 1749-1756 (1994).</li><li>O&#8217;Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microtiter+Dish+Biofilm+Formation+Assay.">Microtiter Dish Biofilm Formation Assay.</a> Journal of Visualized Experience. , (2011).</li><li>Nomoto, M., Ohsawa, M., Wang, H. L., Chen, C. C., Yeh, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+and+Characterization+of+Extracellular+Alkaline+Phosphatase+from+an+Alkalophilic+Bacterium.">Purification and Characterization of Extracellular Alkaline Phosphatase from an Alkalophilic Bacterium.</a> Agricultural and Biological Chemistry. 52 (7), 1643-1647 (1988).</li></ol>

在我们的检测中, 我们使用 pNPP 作为 ALP 底物。这是一个为 ELISA 设计的工作解决方案, 不需要稀释。在 ALP 水解后, 黄色产物会产生, 并可在405纳米处进行测量。在酶反应结束时, 我们对96井板进行了短暂的离心, 并将上清液转移到一个新的96井板上, 用板读取器测量吸收率。我们发现, 这种额外的离心步骤是至关重要的, 因为它增加了在 405 nm 的吸收浓度, 可能是通过消除酶反应过程中产生的任何可能的...

碱性磷酸酶 (ALP) 在原核细胞和真核细胞1中普遍表达。它可以催化不同分子的单磷酸盐水解, 如核苷酸、蛋白质、生物碱、磷酸和酸酐。在人类中, 真核 ALP 存在于许多组织中, 包括肝脏、骨骼、肠道和胎盘2。它在蛋白质磷酸化、细胞生长、凋亡、干细胞过程以及正常的骨骼矿化中起着重要作用。真核 alp 也是骨、肝脏和其他组织器官升高时存在疾病的关键血清指标.
原核 alp 已在各种细菌细胞中检测到, 包括大肠杆菌 5、金黄色葡萄球菌6、7和土壤中一些常见的瘤胃细菌8。细菌 ALP 活性已被用作检测农药、重金属9和细菌污染10的生物传感器。利用 ALP 的本构表达法鉴定葡萄球菌11 和区分沙雷氏菌和肠杆菌12。进一步指出,本构 alp 的产生与葡萄球菌13的致病性有关。虽然 alp 在不同的环境中已经被研究了3,4, 但很少报告其在生物膜培养中的活动和功能。
据记录, 与自由生活的细菌细胞对照14相比, 生物膜具有不同的细菌寿命.在金黄色葡萄球菌中, 生物膜的形成已被确定在各种临床条件下, 并解释了抗生素耐药和慢性炎症 15,16。据报道, 许多分子在生物膜基质中被发现, 如多糖、蛋白质、核酸和脂类, 但生物膜的形成机制尚不完全清楚 14.为了了解 ALP 在生物膜形成中的作用, 我们在96个井组织培养板中培养了金黄色葡萄球菌生物膜, 并使用对硝基苯磷酸酯 (pNPP) 测定了 alp 活性。
分子 pNPP 是 alp 的现成基板, 已被广泛用于测量 alp 活性6,17, 18.这种比色测定的基础是将对硝基苯磷酸酯 (pNPP) 转化为对硝基酚, 使有色产品的含量达到405纳米。与其他传统的 ALP 检测方法相比, 如琼脂糖凝胶电泳19、小麦胚芽凝集素 (wga) 沉淀和 WGA-HPLC20, 该检测方法具有高度的特异性、敏感性、易于繁殖性, 最重要的是, 允许高吞吐量。

<table>
	<tbody>
		<tr>
			<td>Agar</td>
			<td>VWR</td>
			<td>9002-18-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf Centrifuge</td>
			<td>Thomas Scientific</td>
			<td>5810</td>
			<td></td>
		</tr>
		<tr>
			<td>Gluose</td>
			<td>VWR</td>
			<td>50-99-7</td>
			<td></td>
		</tr>
		<tr>
			<td>NaOH pellets</td>
			<td>VWR</td>
			<td>SS0550-500GR</td>
			<td></td>
		</tr>
		<tr>
			<td>Para-nitrophenylphosphate (pNPP)&#160;</td>
			<td>Sigma</td>
			<td>P7998-100ML</td>
			<td>Typical concentrations of pNPP liquid substrates, often used in enzyme-linked immunesorbent assays (ELISA), range between 10 to 50 mM. Similar to most ready-to-use pNPP liquid substrates like the one used here, the exact pNPP concentration is not disclosed due to its proprietary nature.</td>
		</tr>
		<tr>
			<td>10X PBS, pH7.4. 
			173 mM NaCl, 
			2.7 mM KCl, 
			8 mM Na2HPO4, 
			2 mM KH2PO4</td>
			<td>Sigma</td>
			<td>P3288-1VL</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate Reader</td>
			<td>Biotek</td>
			<td>ELx808</td>
			<td></td>
		</tr>
		<tr>
			<td>S. aureus</td>
			<td>ATCC</td>
			<td>ATCC25923</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptic Soy Agar&#160;&#160;&#160;&#160;&#160;&#160; 15g / L TSB</td>
			<td>VWR</td>
			<td>9002-18-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptic Soy Broth:&#160;&#160;&#160;&#160;&#160; g/L 
			Pancreatic Digest of Casein............ 15.0 
			Papaic Digest of Soyben Meal.........5.00 
			Sodium chloride.............................. 3.00 
			Dextrose........................................&#160; 2.50 
			Dipotassium phosphate..................2.50</td>
			<td>VWR</td>
			<td>90006-098</td>
			<td></td>
		</tr>
		<tr>
			<td>96 well tissue culture plates</td>
			<td>BD</td>
			<td>6902D09</td>
			<td>U shaped bottom</td>
		</tr>
	</tbody>
</table>

colorimetric analysis of alkaline phosphatase activity in s. aureus biofilm

碱性磷酸酶 (ALP) 是一种常见的酶, 表达在原核细胞和真核细胞中。它催化了在碱性 ph 值下从许多分子中水解的一酯磷酸酯, 并在磷酸盐代谢中起着不可或缺的作用。在人类中, 真核 ALP 是诊断各种疾病 (如胆汁淤积和软骨病) 最常用的酶信号之一。在s.金黄色葡萄球菌, alp 是完全在细胞膜上检测到的;它也被表达为一个分泌形式, 以及。然而, 人们对它在生物膜形成中的作用了解甚少。
这份手稿的目的是开发一种快速可靠的检测方法, 以测量不需要蛋白质分离的金黄色葡萄球菌生物膜中的 alp 活性。以对硝基苯磷 (pNPP) 为底物, 测定了96孔组织培养板中形成的金黄色葡萄球菌生物膜中的 alp 活性。活性是基于可溶性反应产物的形成测量 405 nm 吸收。96井组织培养板方法的高吞吐量特性为 ALP 活性检测提供了一种灵敏、可重复的方法。同样的实验设置也可以扩展到测量与生物膜形成有关的其他细胞外分子标记。

图 1显示了96种井组织培养板中金黄色葡萄球菌生物膜培养的 alp 活性的代表性结果。在每口井中加入75μl 的商用 pNPP 溶液, 并在室温下孵育。孵育不同时间 (0分钟、15分钟、30分钟、45分钟、60分钟、、75分钟) 后, 分别加入 75μl 5M NaOH 以阻止反应。然后在 405 nm 的96井板读取器中测量了该产品。每个时间点都在一个96孔板的三胞胎中重复, 整个实验?...

Watch this Scientific Journal Video about Colorimetric Analysis of Alkaline Phosphatase Activity in S. aureus Biofilm at JoVE.com

Colorimetric Analysis of Alkaline Phosphatase Activity in S. aureus Biofilm

在这篇手稿中, 我们建立了一个高吞吐量的方法来量化碱性磷酸酶活性的金黄色葡萄球菌生物膜培养96孔组织培养板

金黄色葡萄球菌生物膜碱性磷酸酶活性的比色法分析

Alkaline phosphatase (ALP) is a common enzyme expressed in both prokaryotic and eukaryotic cells. It catalyzes the hydrolysis of phosphate monoesters from ...

该协议提出了一种简单可靠的方法，用于测量S.aureus生物膜培养物中的碱化磷酸酶活性。这将有助于我们了解ALP在生物膜形成中的功能。这种技术的主要优点是高吞吐量。它非常敏感，易于执行。演示这个程序的将是凯文·达尼科夫斯基，一个前哈珀学院的学生。要准备TSB，在蒸馏水中加入甲壳素胰腺消化、大豆粉的木瓜消化、氯化钠、葡萄糖和磷酸二钾。用铝箔盖住烧瓶，并放入自给器中，在使用前进行消毒。要准备 TSA，请将 agar 添加到 TSB 中，并将其放入自动保护器中进行灭菌。然后让它冷却到室温。接下来，以每盘20毫升的比例倒入培养皿中。要准备生物膜培养介质，在自动杀菌TSB中加入葡萄糖，达到每升10克的浓度。使用真空过滤套件，使用 0.2 微米孔径过滤器过滤溶液。要接种，使用灭菌接种循环从培养皿中挑选一个葡萄球菌葡萄球菌的单个菌落，并放在装满10毫升TSB的培养管中。将管子放入孵化器中，在37摄氏度下过夜生长。使用微管，绘制 100 微升的隔夜培养液，并转移到充满 10 毫升 TSB 的培养管中，用葡萄糖稀释 1：100 体积比。轻轻的涡流混合，获得一致的浓度。使用微管将200微升稀释培养液从一个96孔板中输送到总共18口井中。在37摄氏度下孵育96井板，24小时进行生物膜形成。对于S.aureus的最佳生物膜形成，最好选择一个平底或圆形底部96井板。之后，稍微倾斜板，通过从每一个井的边缘的吸气来抽取上经剂。用 1X PBS 清洗每个井。将板放入离心机中，以每分钟8000转的速度旋转5分钟，然后通过吸气将上一代液。将 75 微升缓冲 ALP 基板（由市售 PNPP 组成）添加到每一井中，并使用计时器记录三分球的每个时间点。在每个孵化时间点后，在三口井中每口加入75微升5个氢摩尔钠，以停止反应。以每分钟 800 转的速度将板离心五分钟。使用移液器将 100 微升的上流液转移到新的 96 井板中，用于热量测量。使用 96 井板读取器测量每一个井在 405 纳米的吸光度。使用 30 分钟的时间点井来控制背景噪音。在三个 96 孔板中重复整个实验。该协议开发一种快速可靠的检测方法，以测量S.aureus生物膜中不需要蛋白质分离的ALP活性。ALP 活动的代表性结果显示，其上升趋势长达 75 分钟。75 分钟后，未观察到 ALP 活性增加。建立良好的生物膜文化是非常重要的，可能需要相当多的试验才能掌握。您可以通过执行我们和其他人先前报告的水晶紫罗兰测定来确认生物膜的形成，或者您可以使用人类血浆对板进行预处理，以方便生物膜的形成。开发后，您可以筛选可能干扰生物膜形成的潜在ALP抑制剂。这一结果将为医学领域的生物膜管理提供重要的信息。五摩尔氢氧化钠在接触时可能很危险。处理时一定要穿戴个人防护装备。

Research

JoVE Journal

Bioengineering

Colorimetric Analysis of Alkaline Phosphatase Activity in S. aureus Biofilm

Quantification of Global Diastolic Function by Kinematic Modeling-based Analysis of Transmitral Flow via the Parametrized Diastolic Filling Formalism

全球舒张功能通过静脉流的通过参数化舒张形式主义运动学建模为基础的定量分析

Quantitative cardiac function assessment remains a challenge for physiologists and clinicians.&#160;Although historically invasive methods have comprised ...

科研

生物工程

Protocol for Biofilm Streamer Formation in a Microfluidic Device with Micro-pillars

协议生物膜形成的流光在微流体器件与微柱

Several bacterial species possess the ability to attach to surfaces and colonize them in the form of thin films called biofilms. Biofilms that grow in ...

Real-time Visualization and Analysis of Chondrocyte Injury Due to Mechanical Loading in Fully Intact Murine Cartilage Explants

全完整型小鼠软骨外种机械负荷对软骨细胞损伤的实时可视化与分析

Homeostasis of articular cartilage depends on the viability of resident cells (chondrocytes). Unfortunately, mechanical trauma can induce widespread ...

A Zebrafish Embryo Model for In Vivo Visualization and Intravital Analysis of Biomaterial-associated Staphylococcus aureus Infection

A Zebrafish Embryo Model for In Vivo Visualization and Intravital Analysis of Biomaterial-associated Staphylococcus aureus Infection

一种用于生物技术相关金黄色葡萄球菌感染活体可视化和内膜分析的斑马鱼胚胎模型

Biomaterial-associated infection (BAI) is a major cause of the failure of biomaterials/medical devices. Staphylococcus aureus is one of the major ...

High-resolution Patterned Biofilm Deposition Using pDawn-Ag43

使用 pDawn-Ag43 的高分辨率图案生物膜沉积

Spatial structure and patterning play an important role in bacterial biofilms. Here we demonstrate an accessible method for culturing E. coli biofilms ...

Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels

三维水凝胶中全球细胞基质金属蛋白酶和代谢活性的测定

Three-dimensional (3D) cell culture systems often more closely recapitulate in vivo cellular responses and functions than traditional two-dimensional (2D) ...

Home-Based Monitor for Gait and Activity Analysis

基于家庭的盖特和活动分析监视器

Current outcomes in neuromuscular disorder clinical trials include motor function scales, timed tests, and strength measures performed by trained clinical ...

Substructure Analyzer: A User-Friendly Workflow for Rapid Exploration and Accurate Analysis of Cellular Bodies in Fluorescence Microscopy Images

子结构分析仪：用于荧光显微镜图像中细胞体快速探索和精确分析的用户友好型工作流程

The last decade has been characterized by breakthroughs in fluorescence microscopy techniques illustrated by spatial resolution improvement but also in ...

Electric and Magnetic Field Devices for Stimulation of Biological Tissues

用于刺激生物组织的电动和磁场设备

Electric fields (EFs) and magnetic fields (MFs) have been widely used by tissue engineering to improve cell dynamics such as proliferation, migration, ...