Combined flat-field and frequency filter approach to correcting artifacts of multichannel two-photon microscopy.
Combined flat-field and frequency filter approach to correcting artifacts of multichannel two-photon microscopy.
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DOI：
10.1117/1.jbo.29.1.016007
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发表时间：
2024-01
影响因子：
3.5
通讯作者：
中科院分区：
文献类型：
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作者：
多光子显微镜(MPM)是一种有用的生物医学成像工具，因为它能够以高分辨率探测标记和未标记的深度分辨组织生物标记物。自动MPM瓷砖扫描允许采集整个幻灯片图像，但可能会受到瓷砖拼接伪影的影响，这阻碍了准确的定量数据分析。我们已经研究了使用ImageJ宏和定制的Python代码的后处理伪影校正方法。使用人十二指肠组织的全载玻片MPM自体荧光图像和二次谐波产生图像对这些方法进行了定量和定性比较。通过使用均方根误差、结构相似性指数和图像直方图测量对处理后的图像和未处理的图像进行评估，来评估去除伪影后的图像质量。对定量和定性结果的考虑表明，与独立用于校正平铺扫描MPM图像的平铺伪影的每种方法相比，定制的基于平场的校正和频率滤波处理步骤的组合提供了改进的伪影校正。虽然这些方法保留了一些图像伪影，但这些处理步骤的进一步优化可能导致用于去除这些在大规模MPM成像中普遍存在的这些伪影的计算高效的方法。在保留原始图像信息的情况下去除这些伪影将有助于在研究和临床环境中使用这种成像方式，其中它在收集组织的详细形态和光学属性方面非常有用。
Multiphoton microscopy (MPM) is a useful biomedical imaging tool for its ability to probe labeled and unlabeled depth-resolved tissue biomarkers at high resolution. Automated MPM tile scanning allows for whole-slide image acquisition but can suffer from tile-stitching artifacts that prevent accurate quantitative data analysis. We have investigated postprocessing artifact correction methods using ImageJ macros and custom Python code. Quantitative and qualitative comparisons of these methods were made using whole-slide MPM autofluorescence and second-harmonic generation images of human duodenal tissue. Image quality after artifact removal is assessed by evaluating the processed image and its unprocessed counterpart using the root mean square error, structural similarity index, and image histogram measurements. Consideration of both quantitative and qualitative results suggest that a combination of a custom flat-field-based correction and frequency filtering processing step provide improved artifact correction when compared with each method used independently to correct for tiling artifacts of tile-scan MPM images. While some image artifacts remain with these methods, further optimization of these processing steps may result in computational-efficient methods for removing these artifacts that are ubiquitous in large-scale MPM imaging. Removal of these artifacts with retention of the original image information would facilitate the use of this imaging modality in both research and clinical settings, where it is highly useful in collecting detailed morphologic and optical properties of tissue.
影响因子：
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作者：
Picon A;Terradillos E;Sánchez-Peralta LF;Mattana S;Cicchi R;Blover BJ;Arbide N;Velasco J;Etzezarraga MC;Pavone FS;Garrote E;Saratxaga CL
通讯作者：
Saratxaga CL