Department of Electrical and Electronic Engineering, The University of Hong Kong, Pokfulam Road, Pokfulam, Hong Kong, China.
Department of Mechanical Engineering, The University of Hong Kong, Pokfulam Road, Pokfulam, Hong Kong, China.
Lab Chip. 2016 May 10;16(10):1743-56. doi: 10.1039/c5lc01458a.
Optical imaging is arguably the most effective tool to visualize living cells with high spatiotemporal resolution and in a nearly noninvasive manner. Driven by this capability, state-of-the-art cellular assay techniques have increasingly been adopting optical imaging for classifying different cell types/stages, and thus dissecting the respective cellular functions. However, it is still a daunting task to image and characterize cell-to-cell variability within an enormous and heterogeneous population - an unmet need in single-cell analysis, which is now widely advocated in modern biology and clinical diagnostics. The challenge stems from the fact that current optical imaging technologies still lack the practical speed and sensitivity for measuring thousands to millions of cells down to the single-cell precision. Adopting the wisdom in high-speed fiber-optics communication, optical time-stretch imaging has emerged as a completely new optical imaging concept which is now proven for ultrahigh-throughput optofluidic single-cell imaging, at least 1-2 orders-of-magnitude higher (up to ∼100 000 cells per second) compared to the existing imaging flow cytometers. It also uniquely enables quantification of intrinsic biophysical markers of individual cells - a largely unexploited class of single-cell signatures that is known to be correlated with the overwhelmingly investigated biochemical markers. With the aim of reaching a wider spectrum of experts specializing in cellular assay developments and applications, this paper highlights the essential basics of optical time-stretch imaging, followed by reviewing the recent developments and applications of optofluidic time-stretch imaging. We will also discuss the current challenges of this technology, in terms of providing new insights in basic biology and enriching the clinical diagnostic toolsets.
光学成像是目前最有效的工具之一,它可以高时空分辨率、近乎非侵入的方式对活细胞进行成像。受此功能的推动,最先进的细胞分析技术越来越多地采用光学成像来对不同的细胞类型/阶段进行分类,从而剖析相应的细胞功能。然而,在一个庞大而异质的群体中对细胞间的变异性进行成像和特征分析仍然是一项艰巨的任务——这是单细胞分析中尚未满足的需求,而单细胞分析现在在现代生物学和临床诊断学中得到了广泛的倡导。这一挑战源于这样一个事实,即当前的光学成像技术在测量成千上万甚至上百万个细胞的速度和灵敏度方面仍存在不足,无法达到单细胞的精度。借鉴高速光纤通信的智慧,光时拉伸成像已经成为一种全新的光学成像概念,它已被证明可用于超高通量的光流控单细胞成像,其速度比现有的成像流式细胞仪至少高 1-2 个数量级(高达每秒约 100000 个细胞)。它还独特地实现了对单个细胞固有生物物理标记的量化,这是一类尚未被充分利用的单细胞特征,已知与被广泛研究的生化标记物相关。为了让更多专门从事细胞分析开发和应用的专家能够了解该技术,本文重点介绍了光时拉伸成像的基本原理,随后回顾了光流控时拉伸成像的最新发展和应用。我们还将讨论该技术目前的挑战,包括在提供基础生物学新见解和丰富临床诊断工具集方面的挑战。