Combining motion analysis and microfluidics--a novel approach for detecting whole-animal responses to test substances.
Combining motion analysis and microfluidics--a novel approach for detecting whole-animal responses to test substances.
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DOI：
10.1371/journal.pone.0113235
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Rundle SD
中科院分区：
文献类型：
--
作者：
Rudin-Bitterli TS;Tills O;Spicer JI;Culverhouse PF;Wielhouwer EM;Richardson MK;Rundle SD
斑马鱼胚胎等小型早期生命阶段越来越多地用于评估化合物在体内的生物效应。然而，此类生物体的行为筛选在数据收集（培养技术、药物输送和成像）和数据评估（非常大的数据集）方面都具有挑战性，与体外测定相比，限制了高通量系统的使用。在这里，我们将微流体流通培养系统或 BioWell 板与新颖的运动分析技术（稀疏光流 - SOF）结合起来，然后进行光谱分析（离散傅立叶变换 - DFT），作为此类筛选自动化数据提取和分析的第一步。将复制的斑马鱼胚胎放置在定制成像系统内的 BioWell 板中，并进行化学暴露（1.5% 乙醇）。在暴露之前（30 分钟）、期间（60 分钟）和之后（60 分钟）对胚胎运动进行录像，然后使用 SOF 提取运动数据（旋转角度和胚胎质心的角度变化）。随后使用 DFT 来量化这些时期表现出的运动模式，并使用多维尺度和 ANOSIM 来测试差异。运动分析显示，与治疗前的运动相比，斑马鱼在酒精暴露期的后半段和恢复期的后半段都有显着的运动变化。手动量化甩尾揭示了与使用自动方法检测到的暴露周期之间相同的差异。然而，自动化方法还结合了生物体中可见的其他运动，例如血流和心跳，并且具有更强的能力来识别环境驱动的生物体行为和生理变化。我们建议，结合这些技术可以提供高效、高通量的测定，用于评估整个胚胎对各种药物和化学品的反应。
Small, early life stages, such as zebrafish embryos are increasingly used to assess the biological effects of chemical compounds in vivo. However, behavioural screens of such organisms are challenging in terms of both data collection (culture techniques, drug delivery and imaging) and data evaluation (very large data sets), restricting the use of high throughput systems compared to in vitro assays. Here, we combine the use of a microfluidic flow-through culture system, or BioWell plate, with a novel motion analysis technique, (sparse optic flow - SOF) followed by spectral analysis (discrete Fourier transformation - DFT), as a first step towards automating data extraction and analysis for such screenings. Replicate zebrafish embryos housed in a BioWell plate within a custom-built imaging system were subject to a chemical exposure (1.5% ethanol). Embryo movement was videoed before (30 min), during (60 min) and after (60 min) exposure and SOF was then used to extract data on movement (angles of rotation and angular changes to the centre of mass of embryos). DFT was subsequently used to quantify the movement patterns exhibited during these periods and Multidimensional Scaling and ANOSIM were used to test for differences. Motion analysis revealed that zebrafish had significantly altered movements during both the second half of the alcohol exposure period and also the second half of the recovery period compared to their pre-treatment movements. Manual quantification of tail flicking revealed the same differences between exposure-periods as detected using the automated approach. However, the automated approach also incorporates other movements visible in the organism such as blood flow and heart beat, and has greater power to discern environmentally-driven changes in the behaviour and physiology of organisms. We suggest that combining these technologies could provide a highly efficient, high throughput assay, for assessing whole embryo responses to various drugs and chemicals.
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DOI：
10.1016/j.vascn.2006.04.004
发表时间：
2007-01-01
影响因子：
1.9
作者：
Parng, Chuenlei;Roy, Nicole Marie;McGrath, Patricia
通讯作者：
McGrath, Patricia
影响因子：
3.4
作者：
MacPhail, R. C.;Brooks, J.;Padilla, S.
通讯作者：
Padilla, S.
影响因子：
3.7
作者：
Ali S;Champagne DL;Alia A;Richardson MK
通讯作者：
Richardson MK
DOI：
10.1002/neu.20290
发表时间：
2006-08-01
期刊：
JOURNAL OF NEUROBIOLOGY
影响因子：
--
作者：
Hicks, Caitlin;Sorocco, Debra;Levin, Michael
通讯作者：
Levin, Michael
影响因子：
48
作者：
Pardo-Martin, Carlos;Chang, Tsung-Yao;Koo, Bryan Kyo;Gilleland, Cody L.;Wasserman, Steven C.;Yanik, Mehmet Fatih
通讯作者：
Yanik, Mehmet Fatih