Russell M P, Vohník S, Thomas G J
Division of Cell Biology and Biophysics School of Biological Sciences, University of Missouri-Kansas City 64110-2499, USA.
Biophys J. 1995 Apr;68(4):1607-12. doi: 10.1016/S0006-3495(95)80334-6.
We describe an ultraviolet resonance Raman (UVRR) spectrometer appropriate for structural studies of biological macromolecules and their assemblies. Instrument design includes the following features: a continuous wave, intracavity doubled, ultraviolet laser source for excitation of the Raman spectrum; a rotating cell (or jet source) for presentation of the sample to the laser beam; a Cassegrain optic with f/1.0 aperture for collection of the Raman scattering; a quartz prism dispersing element for rejection of stray light and Rayleigh scattering; a 0.75-m single grating monochromator for dispersion of the Raman scattering; and a liquid-nitrogen-cooled, charge-coupled device for detection of the Raman photons. The performance of this instrument, assessed on the basis of the observed signal-to-noise ratios, the apparent resolution of closely spaced spectral bands, and the wide spectrometer bandpass of 2200 cm-1, is believed superior to previously described UVRR spectrometers of similar design. Performance characteristics of the instrument are demonstrated in UVRR spectra obtained from standard solvents, p-ethylphenol, which serves as a model for the tyrosine side chain, the DNA nucleotide deoxyguanosine-5'-monophosphate, and the human tumor necrosis factor binding protein, which is considered representative of soluble globular proteins.
我们描述了一种适用于生物大分子及其组装体结构研究的紫外共振拉曼（UVRR）光谱仪。仪器设计包括以下特点：用于激发拉曼光谱的连续波、腔内倍频紫外激光源；用于将样品呈现给激光束的旋转池（或喷射源）；用于收集拉曼散射的f/1.0孔径卡塞格伦光学系统；用于排除杂散光和瑞利散射的石英棱镜色散元件；用于拉曼散射色散的0.75米单光栅单色仪；以及用于检测拉曼光子的液氮冷却电荷耦合器件。基于观察到的信噪比、紧密间隔光谱带的表观分辨率以及2200 cm-1的宽光谱仪通带对该仪器的性能进行评估，结果表明其性能优于先前描述的类似设计的UVRR光谱仪。该仪器的性能特征在从标准溶剂、对乙基苯酚（用作酪氨酸侧链的模型）、DNA核苷酸脱氧鸟苷-5'-单磷酸以及人肿瘤坏死因子结合蛋白（被认为是可溶性球状蛋白的代表）获得的UVRR光谱中得到了证明。
