使用彩色空间光干涉显微镜（cSLIM）定量脑组织中的髓鞘
Quantifying myelin in brain tissue using color spatial light interference microscopy (cSLIM)
摘要：
大脑髓鞘形成不足与神经发育迟缓有关，特别是在高危婴儿中，例如出生时相对于胎龄（SGA）较小的婴儿。需要新的方法来进一步研究这种情况。在这里，我们采用彩色空间光干涉显微镜（cSLIM），它使用明场物镜和RGB相机生成与常规明场图像结合的纳米级灵敏度的路径长度图。使用Luxol Fast Blue染色的组织切片，从明场图像中分割髓鞘结构。使用二元掩模，在相应的相位图中定量分析这些部分。我们首先使用的方法来去除组织脂质和验证cSLIM的灵敏度脂质含量。然后，我们将cSLIM应用于脑组织切片。这些样本来自先前的MRI研究，该研究表明，与小于胎龄（SGA）仔猪相比，适合胎龄（AGA）仔猪的内囊髓鞘形成（ICM）增加，并且水解脂肪饮食改善了两者的ICM。在统计分析之前，对样品的身份设盲。
英文摘要：
Deficient myelination of the brain is associated with neurodevelopmental delays, particularly in high-risk infants, such as those born small in relation to their gestational age (SGA). New methods are needed to further study this condition. Here, we employ Color Spatial Light Interference Microscopy (cSLIM), which uses a brightfield objective and RGB camera to generate pathlength-maps with nanoscale sensitivity in conjunction with a regular brightfield image. Using tissue sections stained with Luxol Fast Blue, the myelin structures were segmented from a brightfield image. Using a binary mask, those portions were quantitatively analyzed in the corresponding phase maps. We first used the CLARITY method to remove tissue lipids and validate the sensitivity of cSLIM to lipid content. We then applied cSLIM to brain histology slices. These specimens are from a previous MRI study, which demonstrated that appropriate for gestational age (AGA) piglets have increased internal capsule myelination (ICM) compared to small for gestational age (SGA) piglets and that a hydrolyzed fat diet improved ICM in both. The identity of samples was blinded until after statistical analyses.
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