OPTICAL LASER TRAP FOR BIOLOGICAL CELL STUDIES

荧光光谱和激光散射将用于 量化辐射引起的加热和细胞损伤影响，可能 发生在脂质体和精子细胞的光学显微操作过程中 使用光学镊子。如果引起加热和热效应，可能会 产生光化学和光生物变化，最终可以 导致细胞结构和功能的改变。评估达到的程度 哪种光学捕获是一种非侵入性和非破坏性的过程，以及 光学激光陷阱将首次与一种 荧光探测和前向光散射测量系统， 用于同时禁闭、微操作和光 脂质体测试颗粒、海星和海星的激发/发射研究 人类精子细胞。研究的前提是，作为聚焦的激光 辐射被细胞吸收，它的温度上升，可以 使脂质体的荧光发射光谱发生移位 含温度依赖膜和核荧光的细胞 探测器。脂质体测试细胞要用荧光染料标记； 被光镊子限制；激发和检测荧光；以及 作为光学镊子参数的函数进行的测量， 包括功率、波长和曝光时间。偏离正常细胞 结构和功能将被用作电池发热和损伤的测试 效果。该项目的具体目标包括：整合 捕集和诊断测量能力.制造和 作为模型细胞样品和荧光宿主的脂质体的校准 薄膜探头.光散射分布和温度的测量 依赖的荧光发射光谱作为激光捕获的函数 参数；激光-细胞相互作用的分析模型；以及， 同样的技术在供暖和蓄电池评估中的应用 海星和人类精子的损伤，使用活动率和精子速度， 鞭毛拍频、膜荧光、核荧光 DNA和染色质结构，以及繁殖能力作为衡量 潜在的伤害。这里描述的技术将为以下方面提供基础 了解光钳辐射对脂质体和精子的影响 细胞，作为重要运输工具的生物系统 人体内的药物、酶和遗传物质。

Fluorescence spectroscopy and laser light scattering will be used to quantify the radiation induced heating and cell damage effects that may occur during the optical micromanipulation of liposomes and sperm cells using optical tweezers. Heating and thermal effects, if induced, may produce photochemical and photobiological changes that can ultimately result in altered cell structure and function. To assess the degree to which optical trapping is a non-invasive and non-destructive process, an optical laser trap is to be integrated, for the first time, with a flourescence detection and forward light scattering measurement system, for the simultaneous confinement, micromanipulation, and light excitation/emission study of liposome test particles, and starfish and human sperm cells. The research premise is that, as focused laser radiation is absorbed by a cell, its temperature increases, and can produce a shift in the fluorescence emission spectrum of liposomes and cells containing temperature-dependent membrane and nuclear fluorescent probes. A liposome test cell is to be tagged with a fluorescent dye; confined by an optical tweezer; fluorescence excited and detected; and measurements made as functions of the optical tweezer parameters, including power, wavelength, and exposure time. Departure from normal cell structure and function will be used as assays for cell heating and damage effects. The specific aims for this project include: integration of the trapping and diagnostic measurement capabilities; fabrication and calibration of liposomes as model cell samples and hosts for fluorescent membrane probes; measurement of light scattering profiles and temperature dependent fluorescence emission spectra as a function of laser trapping parameters; analytical modeling of the laser-cell interaction; and, application of the same techniques to the assessment of heating and cell damage in starfish and human sperm, using motility and sperm velocity, flagellar beat frequency, membrane fluorescence, nuclear fluorescence of DNA and chromatin structure, and reproduction capability as a measure of potential damage. The techniques described herein will provide a basis for understanding optical tweezer radiation effects on liposomes and sperm cells, biological systems that function as important transport vehicles for drugs, enzymes, and genetic material within the human body.