Quantifying myelination at the individual axon scale using color spatial light interference microscopy (cSLIM) (Conference Presentation)

G. Popescu
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Abstract

Deficient myelination in the internal capsule of the brain is associated with neurodevelopmental delays, particularly in high-risk infants such as those born small for gestational age (SGA). MRI technology has been effective at measuring brain growth and composition but lacks myelin specificity and is low resolution. There is an unmet need for developing of new quantitative approaches that are rapid and precise, which can complement MRI and provide insight into the pathology of deficient myelination and efficacy of nutritional interventions. To meet this challenge, we developed Color Spatial Light Interference Microscopy (cSLIM), a method that is cable of generating refractive index maps of stained specimens. Using paraffin embedded brain tissue sections, stained myelin was segmented from a brightfield image and, using a binary mask, those portions were quantitatively analyzed by cSLIM. Due to cSLIM’s nanoscale sensitivity to optical pathlengths and independence with respect to the stain intensity, we quantified subtle variations in myelin density at the single axon scale.
利用彩色空间光干涉显微镜(cSLIM)在单个轴突尺度上定量髓鞘形成(会议报告)
脑内囊髓鞘形成缺陷与神经发育迟缓有关,特别是在高危婴儿中,如出生时小于胎龄的婴儿(SGA)。MRI技术在测量大脑生长和组成方面是有效的,但缺乏髓磷脂特异性和低分辨率。目前还没有满足开发新的定量方法的需求,这些方法既快速又精确,可以补充MRI,并提供对髓鞘形成缺陷病理和营养干预效果的见解。为了应对这一挑战,我们开发了彩色空间光干涉显微镜(cSLIM),这是一种生成染色标本折射率图的方法。使用石蜡包埋脑组织切片,从明场图像中分割染色髓磷脂,并使用二值掩膜,用cSLIM对这些部分进行定量分析。由于cSLIM对光路长度的纳米级敏感性和染色强度的独立性,我们量化了单轴突尺度上髓磷脂密度的细微变化。
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