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首页> 外文学位 >Physiological monitoring of optically trapped cells: Studying the effects of confinement by 1064 nm laser-tweezers using microfluorometry.
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Physiological monitoring of optically trapped cells: Studying the effects of confinement by 1064 nm laser-tweezers using microfluorometry.

机译：光学捕获细胞的生理监控：使用微荧光法研究1064 nm激光镊子的限制作用。

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摘要

A novel technique that combines microfluorometric detection and optical laser trapping has been developed for in-situ assessing the physiological state of an optically trapped biological sample. This optical diagnostic technique achieves high sensitivity ({dollar}>{dollar}30 dB signal-to-noise ratio) and high spatial resolution ({dollar}sim{dollar}1 {dollar}mu{dollar}m) over a broad spectral range ({dollar}>{dollar}400 nm). The fluorescence spectra derived from exogenous fluorescent probes, including laurdan, acridine orange, propidium iodide and Snarf, are used to assess the effects of optical confinement with respect to temperature, DNA structure, cell viability, and intracellular pH, respectively. In the latter three cases, fluorescence is excited via a two-photon absorption process, using the cw laser trap itself as the fluorescence excitation source. This enables the cw near infrared laser trapping beam to be used simultaneously as an optical diagnostic probe as well as an optical micromanipulator. Using microfluorometry, a temperature increase of less than several degrees centigrade was measured for test samples, including liposomes, Chinese hamster ovary (CHO) cells and human sperm cells that were held stationary by 1064 nm optical tweezers having a power density of {dollar}sim{dollar}10{dollar}sp7{dollar} W/cm{dollar}sp2.{dollar} Additional physiological monitoring experiments indicated that there is no observable denaturation of DNA, or change of intracellular pH under typical continuous wave laser trapping conditions (P {dollar}le{dollar} 400 mW). Under some circumstances, however, it was possible to achieve a decrease in cell viability with cw trapping, as monitored by a live/dead vital stain. In comparison, significant DNA denaturation and cellular physiological changes (e.g. cell death) were observed when a Q-switched pulsed laser at a threshold of {dollar}sim30mu{dollar}J/pulse was used as trapping source. These results generally support the conclusion that cw laser trapping at 1064 nm wavelength is a safe, non-invasive process and should prove to be of great value for understanding the mechanisms of laser microirradiation effects on living cells held stationary in a near-infrared trapping beam.
机译：已经开发了一种结合了微荧光检测和光学激光捕获的新技术，用于原位评估光学捕获的生物样品的生理状态。这种光学诊断技术可在宽光谱范围内实现高灵敏度（{dollar}> {dollar} 30 dB信噪比）和高空间分辨率（{dollar} sim {dollar} 1 {dollar} mu {dollar} m）范围（{dollar}> {dollar} 400 nm）。来自外源荧光探针（包括劳丹、,啶橙，碘化丙啶和Snarf）的荧光光谱分别用于评估光学限制对温度，DNA结构，细胞活力和细胞内pH的影响。在后三种情况下，使用连续激光阱本身作为荧光激发源，通过双光子吸收过程激发荧光。这使得连续波近红外激光捕获束可以同时用作光学诊断探针和光学显微操纵器。使用微荧光法，测得的测试样品的温度升高不到几摄氏度，其中包括脂质体，中国仓鼠卵巢（CHO）细胞和人类精子细胞，它们被功率密度为{sim}的1064 nm光学镊子保持静止{dollar} 10 {dollar} sp7 {dollar} W / cm {dollar} sp2。{dollar}其他生理监测实验表明，在典型的连续波激光捕获条件下，没有观察到DNA的变性或细胞内pH的变化（P {dol} le {dollar} 400 mW）。但是，在某些情况下，如通过活体/死活体染色监测的那样，使用cw捕集可以实现细胞活力的降低。相比之下，当使用阈值为of30μJ/脉冲的Q开关脉冲激光作为捕获源时，观察到显着的DNA变性和细胞生理变化（例如细胞死亡）。这些结果通常支持以下结论：连续激光在1064 nm波长处捕获是安全的，非侵入性的过程，应该被证明对于理解激光微辐照对在近红外捕获束中保持静止的活细胞的作用机制具有重要价值。 。

著录项

  • 作者

    Liu, Yagang.;

  • 作者单位

    University of California, Irvine.;

  • 授予单位 University of California, Irvine.;
  • 学科 Engineering Biomedical.; Physics Optics.
  • 学位 Ph.D.
  • 年度 1995
  • 页码 41 p.
  • 总页数 41
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物医学工程;光学;
  • 关键词

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