Accurate position tracking of optically trapped live cells
Accurate position tracking of optically trapped live cells
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DOI：
10.1364/boe.5.001026
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发表时间：
2014-04-01
影响因子：
3.4
通讯作者：
Wright, Amanda J.
中科院分区：
文献类型：
--
作者：
McAlinden, Niall;Glass, David G.;Wright, Amanda J.
光阱技术是生命科学研究中的一种强有力的工具，在许多显微镜实验室和设备中越来越普遍。越来越需要直接捕获感兴趣的细胞，而不是将珠粒引入样品，这可能影响样品的基本生物功能并影响用户希望观察和测量的特性。然而，在跟踪大的非均匀物体（例如细胞）时的不稳定性可以使跟踪位置、校准陷阱强度和进行可靠的测量具有挑战性。这些不稳定性通常表现为细胞滚动或重新取向，并且可以由于粘性阻力和热对流而发生，以及由于布朗力而自发发生。在本文中，我们讨论和数学模型的原因，这卷，并提出了几个实验方法来解决这些问题，包括使用一种新的光束轮廓组成的三个紧密间隔的陷阱和跟踪被困对象通过分析荧光图像。本文提出的方法捕获形成适应性免疫反应系统的一部分的T细胞，但原则上可以应用于广泛的样本，其中捕获对象的大小和不均匀性可能会阻碍粒子跟踪实验。(C)2014年美国光学学会
Optical trapping is a powerful tool in Life Science research and is becoming common place in many microscopy laboratories and facilities. There is a growing need to directly trap the cells of interest rather than introduce beads to the sample that can affect the fundamental biological functions of the sample and impact on the very properties the user wishes to observe and measure. However, instabilities while tracking large inhomogeneous objects, such as cells, can make tracking position, calibrating trap strength and making reliable measurements challenging. These instabilities often manifest themselves as cell roll or re-orientation and can occur as a result of viscous drag forces and thermal convection, as well as spontaneously due to Brownian forces. In this paper we discuss and mathematically model the cause of this roll and present several experimental approaches for tackling these issues, including using a novel beam profile consisting of three closely spaced traps and tracking a trapped object by analysing fluorescence images. The approaches presented here trap T cells which form part of the adaptive immune response system, but in principle can be applied to a wide range of samples where the size and inhomogeneous nature of the trapped object can hinder particle tracking experiments. (C) 2014 Optical Society of America