IDBR: Optical Fourier Processing Microscope Based on Two-Dimensional Gabor Filters

基本的生物过程，如细胞程序性死亡、细胞分化和细胞分裂，涉及亚细胞单位（细胞器）结构的时间依赖性动态变化。这些结构变化是由重要的分子途径控制的，必须量化才能更全面地了解细胞功能。本项目开发的显微仪器将快速成像和量化，具有纳米级灵敏度，细胞器动力学。该方法利用由首席研究员开发的暗场显微镜方法，通过自动探测和定量局部样品纹理，以非常高的灵敏度和在大样本区域上快速绘制亚细胞结构变化。因此，该技术适用于基础生物学研究或药物发现的未染色生物样品的快速筛选。作为常规显微观察的一部分，该仪器还可能通过对动态细胞器变化的高灵敏度测量来揭示新的亚细胞动力学。仪器收集的数据可以与分子成像数据相结合，以提供活细胞内细胞器功能的多模态研究。该仪器对生物学研究的有用性将通过细胞凋亡过程中线粒体裂变的测量来证明。人们越来越认识到线粒体在许多细胞过程中发挥着重要作用，包括癌症等疾病。因此，可以追踪线粒体形状变化的结构标记物可能会对许多新兴的生物学研究产生广泛的影响。该仪器开发项目举例说明了工程设计在方法中普遍存在的原则，这些方法能够测量、理解和控制生物实体，并将用于各种教育和培训活动，涉及博士后、研究生和本科生水平的学生，以及来自当地公立中学和高中的代表性不足的少数民族。

Fundamental biological processes, such as programmed cell death, cell differentiation and cell division, involve time-dependent dynamic alterations in the structure of subcellular units, the organelles. These structural changes are controlled by important molecular pathways, and must be quantified to gain a more complete understanding of cellular function. The microscopic instrument developed in this project will rapidly image and quantify, with nano-metric sensitivity, organelle dynamics. The method utilizes a dark-field microscopic method developed by the Principal Investigator, and allows rapid mapping of subcellular structural changes by automatically probing and quantifying local sample texture with very high sensitivity, and over large sample areas. The technique is therefore applicable to rapid screening of unstained biological samples for basic biological research or drug discovery. The instrument is also likely to reveal novel subcellular dynamics by enabling highly sensitive measurement of dynamic organelle changes as part of routine microscopic observation. Data collected with the instrument may be combined with molecular imaging data to provide a multimodal study of organelle function within living cells. The usefulness of the instrument to biological research will be demonstrated by measurements of mitochondrial fission during apoptosis. Mitochondria are being increasingly recognized as significant players in a number of cellular processes, including diseases such as cancer. As such, structural markers that can track changes in mitochondrial shape due to the deregulation of fusion and fission are likely to have a wide-raging impact on a number of emerging biological studies. This instrument development project exemplifies the principle that engineering design is prevalent in methods, which enable measurement, understanding, and control of biological entities, and will be used in various educational and training activities involving students at the postdoctoral, graduate, and undergraduate levels, as well as under-represented minorities from local public middle schools and high schools.