Adaptive interferometric light-sheets for resolution enhanced imaging with and without labeling

自适应干涉光片，用于带或不带标记的分辨率增强成像

• 批准号: 269858105
• 负责人: Professor Dr. Alexander Rohrbach
• 金额: --
• 依托单位: Institut für Mikrosystemtechnik (IMTEK)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/269858105?language=en
• 关键词: Adaptive; interferometric; light; sheets; resolution

Understanding light matter interaction is of significant importance in modern imaging systems. This is the more the case, the larger and stronger scattering the object is that need to be investigated. Light propagation through weekly absorbing matter primarily changes the phase of light. Therefore phase correcting, adaptive optics has been used in astronomy for quite a while, but recently also in modern optical microscopy. Hence, also in light sheet based microscopy spatial light modulators are used to modulate the phase and the intensity of illumination beams, enabling in principle many advantages to 3-D image acquisition. In this context, Bessel beams with their conical phase profile reveal an amazing capability of beam self-reconstruction and a penetration depth which is increased by about 50% relative to conventional Gaussian beams. However, around their narrow main intensity peak, Bessel beams carry a concentric ring system, which results in a loss of image contrast in light sheet microscopy, if no special tricks are applied. In this proposal we want to use linear and nonlinear optical methods to improve the quality of illumination beams in light sheet microscopy by investigating the dependency of computer- holographically generated phase profiles on the beam propagation properties. By using nonlinear optical concepts, we aim to improve the quality of single laser beams and thereby of the illuminating light sheet, such that the influence of the Bessel beams ring system is nonlinearly suppressed. On the one hand we will use the principle of two-photon fluorescence excitation, where especially the influence of the holographically shaped phase on the propagation of short laser pulses through the scattering medium is to be investigated. On the other hand, we will apply the STED-principle, where we will use a second self-reconstructing Bessel beam of higher order to deplete the fluorescence in the ring system by stimulated emission. Here, we want to minimize the thickness of the light sheet and thereby to maximize the optical resolution by improving the depletion efficiency. In a second step, we want to optimize the phase profiles of the excitation beam and the STED- beam by a feedback holographic control, such that the fluorescence generated in the single beams will be improved significantly and thereby the quality of the light sheet.

在现代成像系统中，了解光质相互作用至关重要。情况越多，需要研究的物体越大越强的散射。通过每周吸收物质的光传播主要改变了光的阶段。因此，相校正，自适应光学已经在天文学中使用了一段时间，但最近也用于现代光学显微镜。因此，在基于光板的显微镜空间光调节器中，也用于调节照明束的相位和强度，从而使3-D图像采集具有许多优势。在这种情况下，贝塞尔梁具有其圆锥形相位剖面，揭示了光束自我重建和穿透深度的惊人能力，相对于传统的高斯梁而言，它增加了约50％。然而，贝塞尔束周围的狭窄主强度峰携带一个同心环系统，如果没有使用特殊技巧，则在光板显微镜中导致图像对比度的损失。在此提案中，我们希望使用线性和非线性光学方法来通过研究计算机全文产生的相位剖面对光束传播特性的相位依赖性来提高光板显微镜中的照明束的质量。通过使用非线性光学概念，我们旨在提高单个激光束的质量，从而提高照明光板的质量，从而非线性地抑制贝塞尔梁环系统的影响。一方面，我们将使用两光子荧光激发的原理，尤其是在该原理中，应研究全息相对短激光脉冲通过散射介质传播的影响。另一方面，我们将应用Sted-principle，在这里我们将使用第二个自我重建的贝塞尔梁高阶的贝塞尔束来通过刺激的发射来耗尽环系统中的荧光。在这里，我们希望最大程度地减少光板的厚度，从而通过提高耗竭效率来最大化光学分辨率。在第二步中，我们希望通过反馈全息控制来优化激发梁和steD梁的相位曲线，以便将单光束中产生的荧光显着改善，从而显着改善光板的质量。