将固定的组织样本用X-CLARITY™水凝胶溶液试剂盒进行孵育,以使水凝胶单体在样品中均匀扩散,然后将其置于X-CLARITY™聚合系统中。单体在厌氧环境中聚合,将生物分子连接到坚固的水凝胶网络,保存分子信息和结构完整性。组织-水凝胶混合物在X-CLARITY™组织清除系统II中使用现成的电泳组织清除溶液。脂质可以通过电泳或被动提取,留下稳定透明的组织-水凝胶混合物,可以通过化学方法进行分子表型分析。
然后使用DeepLabel™抗体染色试剂盒对透明样品进行抗体标记,以增强抗体渗透到透明组织。DeepLabel™ 针对透明的组织样本进行了优化,可用于通过各种清除方法清除的组织,如被动或主动 CLARITY、iDISCO、3DISCO、UDISCO、Visikol 或 CUBICO。
成像前,将组织-水凝胶混合物置于折射率(RI)匹配溶液中,以便使组织内的环境与溶液均匀化。这减少了光线散射,从而增加了光学透明度,则增加了图像质量和成像深度。
“[The X-CLARITY] has actually been fundamental to the Data Brain Project because without being able to clear tissues of the lipids, we wouldn’t be able to image deep into brains. So we wouldn’t be able to extract that structural information that is so important to understand the function of mammalian brains. It’s been a huge advantage and a great leap forward in imaging.”
“Thanks to X-CLARITY, we now have access to the threedimensions of the whole heart organ.”