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arXiv 2003.05851physics.opticsphysics.bio-ph
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用于多焦平面显微镜的自适应光学

Adaptive optics for multifocal plane microscopy

Benjamin Gore, Noah Schwartz, Paul Dalgarno

摘要:

多焦平面显微镜(MUM)允许三维物体在单个相机帧中成像。我们的方法使用双正交衍射相位光栅与二次失真的线应用散焦的第一衍射级,当与中继透镜配对时,允许9个焦平面被成像在一个单一的相机芯片。这种方法需要强信号电平以确保衍射光中的足够强度,但具有紧凑且易于实现的优点。当显微镜开始聚焦到样品更深处时,样品介质中折射率失配和不均匀性引起的像差对信号质量产生不利影响。本文研究了将自适应光学(AO)应用于多焦平面显微镜的成像质量改善。单个校正装置(8x8可变形反射镜(DM))与基于图像的AO控制策略组合以执行光学像差的校正。我们比较完整的端到端的建模结果,使用一个既定的数值建模系统,适用于显微镜实验室结果的测试样品和一些生物样品。最后,我们将证明,结合AO和MUM,我们能够提高生物样本的图像质量,并提供良好的校正整个生物样本的体积。

英文摘要:

Multifocal plane microscopy (MUM) allows three dimensional objects to be imaged in a single camera frame. Our approach uses dual orthogonal diffraction phase gratings with a quadratic distortion of the lines to apply defocus to the first diffraction orders which, when paired with a relay lens, allows for 9 focal planes to be imaged on a single camera chip. This approach requires a strong signal level to ensure sufficient intensity in the diffracted light, but has the advantage of being compact and straightforward to implement. As the microscope begins to focus deeper into the sample, aberrations caused by refractive index mismatch and inhomogeneity in the sample's media have an adverse effect on the signal's quality. In this paper, we investigate the image quality improvement brought by applying adaptive optics (AO) to multifocal plane microscopy. A single correction device (an 8x8 deformable mirror (DM)) is combined with an image-based AO control strategy to perform the correction of optical aberrations. We compare full end-to-end modelling results using an established numerical modelling system adapted for microscopy to laboratory results both on a test sample and on a number of biological samples. Finally, we will demonstrate that combining AO and MUM, we are able to improve the image quality of biological samples and provide a good correction throughout the volume of the biological sample.

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