YIELD SINGLE MOLECULE IN FCS COLLECTION VOLUME BY NUMERICAL SIMULATION

通过数值模拟获得 FCS 收集体积中的单分子

• 批准号: 6206145
• 负责人: SAMUEL T HESS
• 金额: $ 0.04万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6206145
• 关键词: animal tissue; bioengineering /biomedical engineering; biological products; biomedical equipment development; biomedical resource; connective tissue; human tissue; lasers; nervous system; technology /technique

We are investigating the uses of multiphoton autofluorescence imaging and spectroscopy in tissue for basic biological research and as a potential diagnostic tool for pathologists. Intrinsic tissue fluorescence can potentially provide significant information about the structural and chemical composition of tissue. Multiphoton excitation has an increased potential for excitation of intrinsic fluorophores since emission separation can be very efficient (excitation is far removed from the emission region) and the technique possesses an intrinsic optical sectioning important in scattering tissues. When performing multiphoton microscopy with excitation in the 690-750nm region, tissue autofluorescence becomes prevalent at specimen powers of 10-15mW (~3-fold higher powers than those used for 2PM of conventional fluorophores). The clear advantage of this method is that the tissue is potentially free from any fixation or staining artifacts. Dye penetration through a thick sample is often poor and fixation and dehydration techniques are associated with an ~80% shrinkage, a collapse in which structural relationships are not necessarily preserved. The disadvantage of utilizing autofluorescence is that natural fluorophores are clearly not optimized for their fluorescent properties. Optimization of instrumentation is thus essential for the acquisition of good autofluorescence images.

我们正在研究多光子自发荧光的应用 用于基础生物学研究的组织成像和光谱学， 作为病理学家的潜在诊断工具。 固有组织 荧光可以潜在地提供关于 组织结构和化学组成。 多光子激发 具有增加的激发内在荧光团的潜力 由于发射分离可以非常有效（激发远 从发射区移除），并且该技术具有 在散射组织中重要的内在光学切片。 当 在690- 750 nm的激发下进行多光子显微术， 区域，组织自发荧光在样本功率下变得普遍 10- 15 mW（功率比2PM高约3倍， 常规荧光团）。 这种方法的明显优点是 组织可能没有任何固定或染色 藏物 染料透过厚样品的渗透性通常很差 固定和脱水技术与约80%的 收缩，一种结构关系不再 必须保存。 利用自体荧光的缺点 天然的荧光团显然不是最佳的， 荧光特性 因此，仪器的优化是 这对于获得良好的自体荧光图像至关重要。