Abstract:
Three-dimensional (3D) cell culture models such as spheroids and organoids are widely used in the field of experimental biology. To analyze these 3D experimental models, formalin-fixed paraffin-embedded (FFPE) sections are superior to whole-mount imaging for some experimental purposes, such as exploring samples with a depth limitation of primary antibody penetration immunohistochemically. However, tiny 3D cell culture samples are difficult to embed in paraffin and acquire appropriate sections. In this report, we optimized a protocol of paraffin embedding for spheroids and organoids. In addition, we compared FFPE sections with frozen sections in ratio of sample collection and section condition after staining, and could reproduce improved results reliably. The protocol we established could be widely used in many laboratories and become a useful technique for analyzing spheroids and organoids.
摘要:
三维(3D)细胞培养模型如球体和类器官被广泛应用于实验生物学领域。为了分析这些3D实验模型,福尔马林固定石蜡包埋(FFPE)切片在某些实验目的优于整体安装成像,例如用免疫组织化学方法探索具有一级抗体穿透深度限制的样品。然而,微小的3D细胞培养样品很难嵌入石蜡中并获得适当的切片。在这份报告中,我们优化了球体和类器官的石蜡包埋方案。此外,我们比较了FFPE切片和冷冻切片的样品采集比例和染色后的切片条件,并且可以可靠地重现改进的结果。我们建立的协议可以在许多实验室中广泛使用,并成为分析球体和类器官的有用技术。