Ojeda Jenifer F, Xie Changan, Li Yong-Qing, Bertrand Fred E, Wiley John, McConnell Thomas J
Opt Express. 2006 Jun 12;14(12):5385-93. doi: 10.1364/oe.14.005385.
The ability to identify specific chromosomes with certainty has been established by the development of several cytogenetic techniques based on staining. Here, we report the use of a new optical technique, laser tweezers and Raman spectroscopy (LTRS), to capture and manipulate chromosomes in order to obtain their spectral patterns for molecular analysis without the need for staining. The purpose of this study was to obtain Raman spectroscopy patterns for chromosomes number 1, 2, and 3 and to test if the Raman spectroscopy pattern could be used to distinguish these three chromosomes. In our experiment, optical tweezers were used to capture the individual chromosomes and the Raman spectral patterns were collected for the trapped chromosomes. Then, the captured chromosome was manipulated with the optical tweezers and moved to another chamber through a micro - channel, in which the chromosomes were G- banded for positive identification as chromosome number 1, 2, or 3. Generalized discriminate analysis (GDA) was used to compare the Raman signatures. This analysis revealed that chromosomes 1, 2, and 3 could be distinguished and identified based on their Raman spectra. Development of this approach will lead to more rapid automatic methods for chromosome analysis and identification without the use of prior staining. Moreover, the Raman spectral patterns may lend themselves to more detailed analysis of chromosomal structure than is currently available with standard staining protocols. Such analysis may some day be useful for rapid, automated screening and diagnosis for certain cancers.
通过基于染色的几种细胞遗传学技术的发展，已经确立了准确识别特定染色体的能力。在此，我们报告了一种新的光学技术——激光镊子与拉曼光谱技术（LTRS）的应用，该技术用于捕获和操纵染色体，以便在无需染色的情况下获取其光谱模式用于分子分析。本研究的目的是获得1号、2号和3号染色体的拉曼光谱模式，并测试拉曼光谱模式是否可用于区分这三条染色体。在我们的实验中，使用光镊捕获单个染色体，并收集被捕获染色体的拉曼光谱模式。然后，用光学镊子操纵捕获的染色体，并通过微通道将其移动到另一个腔室，在该腔室中对染色体进行G带染色以阳性鉴定为1号、2号或3号染色体。使用广义判别分析（GDA）来比较拉曼特征。该分析表明，1号、2号和3号染色体可以根据它们的拉曼光谱进行区分和识别。这种方法的发展将导致在不使用预先染色的情况下实现更快速的染色体分析和识别自动化方法。此外，与目前的标准染色方案相比，拉曼光谱模式可能更有助于对染色体结构进行更详细的分析。这种分析有朝一日可能对某些癌症的快速、自动化筛查和诊断有用。
