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.

生物组织体内研究的进展本质上依赖于空间和时间分辨率不断提高的显微镜。在本提案中，我们通过使用新型自适应光学元件和控制技术来提高共焦激光扫描显微镜的空间分辨率来应对这一挑战。为了产生三维图像，共焦显微镜通常通过移动光学元件并使用压电镜或检流镜来使用聚焦激光束扫描体积，其中速度受到质量和惯性的限制。这可能进一步导致运动伪影并且需要庞大的设置，从而限制了小型化的潜力。原则上，使用具有高数值孔径的显微镜物镜可以最小化光斑尺寸，从而最小化分辨率。然而，由于它们针对具有有限视场的单个焦平面进行了优化，因此除了样品引起的像差之外，扫描还会导致显着的系统引起的像差，从而限制了可实现的分辨率。为了克服这些问题，可以使用自适应元件进行像差校正和无运动扫描。我们的目标是开发一种自适应共焦显微镜，理想情况下仅使用两个（一个镜头和一个棱镜）自适应光学元件来提供快速三维扫描和实时像差校正，从而在整个视场上进行衍射限制成像。为此，我们将开发新型自适应镜头，结合轴向扫描和像差校正，包括几何像差（散焦、球面、像散、慧差）和色差。同样，我们将使用新型双轴消色差自适应棱镜进行横向扫描。与使用振镜扫描相比，这可以实现更紧凑的共线几何形状。利用这些自适应光学元件，这种新型智能显微镜在小型化和开发稳健的手持系统方面具有巨大的潜力。我们将通过研究致甲状腺肿素对斑马鱼胚胎的影响来展示潜在的应用和增强的成像性能。