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A concept for single-shot volumetric fluorescence imaging via orthogonally polarized excitation lattices

机译:通过正交偏振激发晶格进行单次体积荧光成像的概念

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摘要

The deconvolution of widefield fluorescence images provides only guesses of spatial frequency information along the optical axis due to the so called missing cone in the optical transfer function. Retaining the single-shot imaging speed of deconvolution microscopy while gaining access to missing cone information is thus highly desirable for microscopy of volumetric samples. Here, we present a concept that superimposes two orthogonally polarized excitation lattices with a phase-shift of p between them. In conjunction with a non-iterative image reconstruction algorithm this permits the restoration of missing cone information. We show how fluorescence anisotropy could be used as a method to encode and decode the patterns simultaneously and develop a rigorous theoretical framework for the method. Through in-silico experiments and imaging of fixed biological cells on a structured illumination microscope that emulates the proposed setup we validate the feasibility of the method.
机译:由于光学传递函数中所谓的缺失锥,宽视野荧光图像的去卷积仅提供沿光轴的空间频率信息的猜测。因此,对于体积样品的显微术,非常需要保持解卷积显微镜的单次成像速度,同时获得丢失的锥体信息。在这里,我们提出了一个概念,该概念叠加了两个正交极化的激发晶格,它们之间具有p的相移。结合非迭代图像重建算法,可以恢复丢失的视锥信息。我们展示了如何使用荧光各向异性作为同时编码和解码图案的方法,并为该方法建立严格的理论框架。通过计算机模拟实验以及在模拟拟议设置的结构化照明显微镜上对固定生物细胞进行成像,我们验证了该方法的可行性。

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