Design and implementation of algorithms for focus automation in digital imaging time-lapse microscopy
Design and implementation of algorithms for focus automation in digital imaging time-lapse microscopy
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DOI：
10.1002/cyto.10174
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发表时间：
2002-12-01
期刊：
影响因子：
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通讯作者：
Kron, SJ
中科院分区：
文献类型：
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作者：
LeSage, AJ;Kron, SJ
背景资料：使用Nomarski-DIC的数字延时显微镜要求自动聚焦系统适应细胞形状，大小和位置的变化，同时与显微镜和成像仪中的漂移，噪声和滞后相竞争。我们设计并实现了一个自动对焦系统，该系统可以在动态条件下跟踪对象，并将焦点保持在可分辨的阈值内。Methods：使用Nomarski中经过验证的新颖自动对焦算法，我们对记录的图像堆栈进行“虚拟”实验，以模拟漂移和突然位移，并测试搜索算法的响应。我们发现，将简单的[ 1，-1]对比度函数与自适应的“暖-冷”聚焦算法相结合，在聚焦精度和噪声容限之间产生了合理的折衷。这种方法被实施，以记录酵母细胞的生长动力学在单个和多个领域的views.Conclusions：我们已经实现了一个强大的数字自动对焦，保持专注于光学复杂的样品成像在高分辨率。这个系统的漂移和振动的容忍度表明，它是一个实用的系统，在许多生物应用的时间推移成像。(C)2002 Wiley-Liss，Inc.
Background: Digital time-lapse microscopy using Nomarski-DIC requires that an autofocusing system adapt to changes in cell shape, size, and position while contending with drift, noise, and hysteresis in the microscope and imager. We have designed and implemented an autofocusing system that tracks subjects under dynamic conditions and maintains focus within a threshold of discriminability.Methods: With the use of proven and novel algorithms for autofocusing in Nomarski, we performed "virtual" experiments on recorded image stacks to simulate drift and sudden displacements and test the search algorithm response.Results: We found that combining a simple [ 1, -1] contrast function with an adaptive "warmer- colder" focusing algorithm yields a reasonable compromise between focusing precision and noise tolerance. This method was implemented to record growth kinetics of yeast cells in single and multiple fields of view over several hours.Conclusions: We have implemented a robust digital auto-focus that maintains focus on optically complex samples imaged at high resolution. The tolerance of this system of drift and vibration suggests that it is a practical system for time-lapse imaging in many biological applications. (C) 2002 Wiley-Liss, Inc.