Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay.
Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay.
复制标题
DOI：
10.1002/biot.201400494
复制
发表时间：
2015-03
影响因子：
4.7
通讯作者：
Wadso, Ingemar
中科院分区：
文献类型：
--
作者：
Braissant, Olivier;Keiser, Jennifer;Meister, Isabel;Bachmann, Alexander;Wirz, Dieter;Goepfert, Beat;Bonkat, Gernot;Wadso, Ingemar
等温微量热法是一种无标记的检测方法，可以监测酶和代谢活性。该技术具有优势，但大多数仪器的通量较低，这限制了它们在生物测定中的应用。这里，使用配备有类似于48孔板的容器保持器的等温微量热计。该微量热仪的高通量使其在生物医学和制药应用中具有重要价值。我们的结果表明，该仪器的灵敏度允许检测3 × 104个细菌/小瓶。随着接种量的减少，在2和9小时之间检测到奇异变形杆菌在Luria肉汤培养基中的生长。培养物释放2.1J，最大热功率为76 μW。使用量热和分光光度数据计算的生长速率分别为0.60和0.57 h-1。还可以收集关于奇异变形杆菌的蛋白酶活性与产热的最后一个峰值相匹配的其他见解。还监测了释放最大热功率2.1 μW的肿瘤微组织的生长，这对应于微组织直径从约2.1 μ W增加到约2.1 μ W。100至428 μm。这为癌症研究、诊断和开发新的抗肿瘤药物开辟了新的研究途径。对于寄生虫，该技术允许使用运动和代谢活动评估寄生虫存活，即使是单个蠕虫。
Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 × 104 bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 μW. The growth rate calculated using calorimetric and spectrophotometric data were 0.60 and 0.57 h–1, respectively. Additional insight on protease activities of P. mirabilis matching the last peak in heat production could be gathered as well. Growth of tumor microtissues releasing a maximum thermal power of 2.1 μW was also monitored and corresponds to a diameter increase of the microtissues from ca. 100 to 428 μm. This opens new research avenues in cancer research, diagnostics, and development of new antitumor drugs. For parasitic worms, the technique allows assessment of parasite survival using motor and metabolic activities even with a single worm.
登录
查看更多内容
影响因子：
5.6
作者：
Domenis R;Bisetto E;Rossi D;Comelli M;Mavelli I
通讯作者：
Mavelli I
影响因子：
15.3
作者：
Galindo, FG;Rocculi, P;Sjöhlm, I
通讯作者：
Sjöhlm, I
影响因子：
3.5
作者：
Kabanova, N.;Stulova, I.;Vilu, R.
通讯作者：
Vilu, R.
DOI：
10.1152/ajplung.90445.2008
发表时间：
2009-06-01
影响因子：
4.9
作者：
Ceresa, Claudia C.;Knox, Alan J.;Johnson, Simon R.
通讯作者：
Johnson, Simon R.
影响因子：
1.8
作者：
Hansen, Lee D.;Criddle, Richard S.;Battley, Edwin H.
通讯作者：
Battley, Edwin H.