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用于延时显微镜应用和细胞可视化的低成本运动跟踪系统(LOCOMOTIS)。

Low-cost motility tracking system (LOCOMOTIS) for time-lapse microscopy applications and cell visualisation.

作者信息

Lynch Adam E, Triajianto Junian, Routledge Edwin

机构信息

Institute for the Environment, Brunel University, Uxbridge, London, United Kingdom.

Karang Menjangan, Surabaya, Indonesia.

出版信息

PLoS One. 2014 Aug 14;9(8):e103547. doi: 10.1371/journal.pone.0103547. eCollection 2014.

Abstract

Direct visualisation of cells for the purpose of studying their motility has typically required expensive microscopy equipment. However, recent advances in digital sensors mean that it is now possible to image cells for a fraction of the price of a standard microscope. Along with low-cost imaging there has also been a large increase in the availability of high quality, open-source analysis programs. In this study we describe the development and performance of an expandable cell motility system employing inexpensive, commercially available digital USB microscopes to image various cell types using time-lapse and perform tracking assays in proof-of-concept experiments. With this system we were able to measure and record three separate assays simultaneously on one personal computer using identical microscopes, and obtained tracking results comparable in quality to those from other studies that used standard, more expensive, equipment. The microscopes used in our system were capable of a maximum magnification of 413.6×. Although resolution was lower than that of a standard inverted microscope we found this difference to be indistinguishable at the magnification chosen for cell tracking experiments (206.8×). In preliminary cell culture experiments using our system, velocities (mean µm/min ± SE) of 0.81 ± 0.01 (Biomphalaria glabrata hemocytes on uncoated plates), 1.17 ± 0.004 (MDA-MB-231 breast cancer cells), 1.24 ± 0.006 (SC5 mouse Sertoli cells) and 2.21 ± 0.01 (B. glabrata hemocytes on Poly-L-Lysine coated plates), were measured and are consistent with previous reports. We believe that this system, coupled with open-source analysis software, demonstrates that higher throughput time-lapse imaging of cells for the purpose of studying motility can be an affordable option for all researchers.

摘要

为了研究细胞运动性而对细胞进行直接可视化通常需要昂贵的显微镜设备。然而,数字传感器的最新进展意味着现在可以用标准显微镜价格的一小部分对细胞进行成像。随着低成本成像技术的出现,高质量的开源分析程序的可用性也大幅增加。在本研究中,我们描述了一种可扩展的细胞运动系统的开发和性能,该系统使用廉价的商用数字USB显微镜,通过延时成像对各种细胞类型进行成像,并在概念验证实验中进行跟踪分析。使用该系统,我们能够在一台个人计算机上使用相同的显微镜同时测量和记录三个独立的分析,并获得了与其他使用标准且更昂贵设备的研究质量相当的跟踪结果。我们系统中使用的显微镜最大放大倍数为413.6倍。虽然分辨率低于标准倒置显微镜,但我们发现在为细胞跟踪实验选择的放大倍数(206.8倍)下,这种差异难以区分。在使用我们系统的初步细胞培养实验中,测量了0.81±0.01(未包被平板上的光滑双脐螺血细胞)、1.17±0.004(MDA-MB-231乳腺癌细胞)、1.24±0.006(SC5小鼠支持细胞)和2.21±0.01(聚-L-赖氨酸包被平板上的光滑双脐螺血细胞)的速度(平均μm/分钟±标准误),这些结果与先前报道一致。我们相信这个系统,结合开源分析软件,表明为了研究运动性而对细胞进行更高通量的延时成像对于所有研究人员来说可以是一个经济实惠的选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/77e4/4133191/3f724ed1dea9/pone.0103547.g001.jpg

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