Aberration correction for real-time measurements in adaptive confocal microscopy

自适应共焦显微镜实时测量的像差校正

• 批准号: 271021903
• 负责人: Professor Dr.-Ing. Jürgen W. Czarske
• 金额: --
• 依托单位: Proffesur für Mikroaktorik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/271021903?language=en
• 关键词: Aberration; correction; real; time; measurements

Progress in the in vivo investigation of biological tissue essentially relies on microscopy with ever increasing spatial and temporal resolution. In the present proposal, we address this challenge by aiming to enhance the spatial resolution confocal laser scanning microscopy using novel adaptive optical elements and control techniques. To produce a three dimensional image, confocal microscopy scans the volume with a focused laser beam, typically by moving optical elements and using piezo or galvo mirrors where the speed is limited by mass and inertia. This may further result in motion artefacts and requires a bulky set-up, limiting the potential for miniaturization. The spot size, and hence the resolution, can - in principle - be minimized using microscope objectives with a high numerical aperture. As they are optimized for a single focal plane with a limited field, however, scanning results in significant system-induced aberrations in addition to the sample-induced aberrations and hence limits the achievable resolution. To overcome these problems, one can use adaptive elements both for aberration correction and for motion-free scanning.Our aim is the development of an adaptive confocal microscope that uses ideally only two (one lens and one prism) adaptive optical elements to provide fast three dimensional scanning and real-time aberration correction for diffraction-limited imaging over the whole field of view. For this purpose, we will develop novel adaptive lenses that combine axial scanning and aberration correction, including geometric (defocus, spherical, astigmatism, coma) and chromatic aberrations. Similarly, we will preform the lateral scans with novel bi-axial achromatic adaptive prisms. In contrast to scanning with galvo mirrors, this allows for a more compact collinear geometry. Using these adaptive optical elements, this novel smart microscope has a great potential for miniaturization and for the development of robust handheld systems. We will demonstrate potential applications and the enhanced imaging performance by investigating the effects of goitrogens in zebrafish embryos.

生物组织的体内研究的进展基本上依赖于显微镜，并且随着空间和时间分辨率的增加。在本提案中，我们通过使用新型的自适应光学元素和控制技术来增强空间分辨率共聚焦激光扫描显微镜来解决这一挑战。为了产生三维图像，共焦显微镜使用聚焦激光束扫描体积，通常是通过移动光学元件以及使用质量和惯性限制速度的压电或加尔沃镜子。这可能会进一步导致运动人工制品，并需要笨重的设置，从而限制了微型化的潜力。因此，原则上可以使用具有高数值光圈的显微镜目标将斑点大小及其分辨率最小化。但是，由于它们针对具有有限场的单个焦平面进行了优化，因此除了样品诱导的畸变外，扫描还会导致系统诱导的畸变，从而限制了可实现的分辨率。为了克服这些问题，人们可以使用自适应元素进行畸变校正和无运动扫描。我们的目标是开发自适应共聚焦显微镜，该显微镜仅使用理想的两个（一个镜头和一个棱镜）自适应光学元件来提供快速的三维扫描和快速的三维式扫描和实时的异常校正，以实现扩散范围的视图，而不是整个视图。为此，我们将开发新型的自适应透镜，结合轴向扫描和畸变校正，包括几何（散焦，球形，散光，昏迷）和色差。同样，我们将使用新型的双轴性适应性棱镜进行横向扫描。与使用Galvo镜子进行扫描相反，这允许更紧凑的共线几何形状。使用这些自适应光学元素，这种新型的智能显微镜具有微型化和强大的手持系统的发展潜力。我们将通过研究山尾胚胎中山羊膜的影响来证明潜在的应用和增强的成像性能。