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Vertically scanned laser sheet microscopy

机译:垂直扫描激光片显微镜

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摘要

Laser sheet microscopy is a widely used imaging technique for imaging the three-dimensional distribution of a fluorescence signal in fixed tissue or small organisms. In laser sheet microscopy, the stripe artifacts caused by high absorption or high scattering structures are very common, greatly affecting image quality. To solve this problem, we report here a two-step procedure which consists of continuously acquiring laser sheet images while vertically displacing the sample, and then using the variational stationary noise remover (VSNR) method to further reduce the remaining stripes. Images from a cleared murine colon acquired with a vertical scan are compared with common stitching procedures demonstrating that vertically scanned light sheet microscopy greatly improves the performance of current light sheet microscopy approaches without the need for complex changes to the imaging setup and allows imaging of elongated samples, extending the field of view in the vertical direction.
机译:激光薄片显微镜是一种广泛使用的成像技术,用于对固定组织或小型生物体中荧光信号的三维分布进行成像。在激光薄片显微镜中,由高吸收或高散射结构引起的条纹伪影非常普遍,极大地影响了图像质量。为了解决这个问题,我们在这里报告了一个两步过程,该过程包括在垂直移动样本的同时连续获取激光薄片图像,然后使用可变平稳噪声消除器(VSNR)方法进一步减少残留条纹。将通过垂直扫描获得的清除的小鼠结肠的图像与常规缝合程序进行比较,证明垂直扫描的光片显微镜极大地改善了当前的光片显微镜方法的性能,而无需对成像设置进行复杂的更改,并且可以对细长样品进行成像,在垂直方向上扩展了视野。

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