LASER RAMAN PROBE OF MOLECULAR STRUCTURE IN LIVING CELLS

提出建立一种激光拉曼显微镜，其能够 以大约一微米的分辨率观察染色体材料。 使用 该装置的拉曼光谱将取自各种染色体， 容易获得的来源。 通过聚焦约1微米或更小的激光束， 在染色体的不同部分和相关的细胞 装置并捕获通过该装置反向散射的拉曼光， 显微镜物镜，建议同时获得经典和 各种染色体成分的共振拉曼光谱，包括 蛋白质和DNA。 特别是拉曼标记带特性 DNA的A、B、C和Z形式以及对 将寻找蛋白质二级结构。 Z标记的存在 基因的某一部分中的条带将提供Z型的证据。 多线 将在固定和非固定状态下检查染色体。 该拉曼 含有Z-DNA结合抗体的多线染色体的光谱将 也可以看看是否有证据可以证明存在的 抗体结合后多线染色体DNA中的Z-构象。 这种方法可以解决Z-DNA是否存在于 多线染色体或者抗体是否能够 在结合时将DNA转化为Z型。 通过收集瞬间的拉曼 活细胞的细胞组分的光谱，建议 确定的类型，程度和持续时间的构象变化， 细胞分裂过程中的DNA和相关蛋白质。 因此，希望 Z-DNA是否是一种天然存在的 遗传物质的结构。 也希望能改善 了解遗传过程的分子基础。 等 理解只能有助于制定一个更合理的方法， 用于治疗疾病的药物疗法的发展， 遗传物质的故障。

It is proposed to build a laser Raman microscope which is capable of observing chromosomal material with about one micron resolution. Using this apparatus Raman spectra will be taken of chromosomes from a variety of easily available sources. By focusing a laser beam of about 1 micron or less on various portions of the chromosome and associated cellular apparatus and capturing the Raman light which is back scattered through the microscope objective, it is proposed to obtain both the classical and the resonance Raman spectra of the various chromosomal components including both protein and DNA. In particular the Raman marker band characteristic of the A, B, C, and Z forms of DNA as well as Raman bands sensitive to protein secondary structure will be sought. The presence of the Z marker bands in a portions of a gene would give evidence for the Z form. Polytene chromosomes will be examined in fixed and non-fixed states. The Raman spectra of polytene chromosomes containing bound antibodies to Z-DNA will also be taken to see if evidence can be found for presence of the Z-conformation in the DNA of polytene chromosomes upon antibody binding. This approach could settle the questions of whether or not Z-DNA exists in polytene chromosomes or whether or not antibodies are capable of pulling the DNA into the Z form upon binding. By collecting instanteous Raman spectra of the cellular components of living cells, it is proposed to determine the type, extent of and duration of conformational changes in the DNA and associated proteins during cell division. Thus, it is hoped to answer the questions as to whether Z-DNA is a naturallly occurring structure in genetic material. It is also hoped to improve the understanding of the molecular basis of the hereditary process. Such understanding can only help to formulate a more rational approach to the development of drug therapy for treatment of diseases which involve malfunction of the genetic material.