Novel Optical Technique for Recovery of Fetal Cells in Maternal Blood

项目摘要 目前基于细胞的产前诊断技术，如羊膜穿刺术、绒毛 采样(CVS)和经皮脐血采样(PUBS)是有创性和现场性的 胎儿丢失的重大风险。利用胎儿细胞循环进行无创性产前诊断 孕妇孕期外周血不会造成危险，因此备受关注。 传给胎儿。尽管有几种胎儿细胞类型已经成为目标，但搜索的重点是 胎儿有核红细胞(FNRBC)。然而，由于fNRBC的浓度非常低 在母体血液中，母体来源的有核红细胞(MNRBC)和 未能找到能够区分fNRBC和mNRBC的广泛适用的标识符 可靠地检测和分离足够数量的活细胞用于临床仍然是一项 巨大的挑战。 胎儿NRBC和母体NRBC在核形态和血红蛋白方面都不同 分子结构。最近，麻省理工学院的首席研究员和他的同事们 发展了生物医学光散射光谱(LSS)，并显示了其独特的能力 在不破坏细胞的情况下，光学探测核形态。我们的团队进一步晋级 LSS，并发展了共焦吸收和散射光谱技术(类) 能够监测小共焦体积的亚细胞器。 这项研究计划的目的有两个。 (1)我们将使用我们实验室开发的现有一流设备来鉴定fNRBC 天然光学生物标志物。 (2)基于这些生物标志物，我们将开发一种快速、简单和健壮的光学技术 它在同一仪器中结合了光学显微镜和光学镊子，它将 能够从孕妇外周血样本中浓缩和回收完整的fNRBC。 这将导致一种临床上有用的产前基因检测方法。

Project Summary Present cell-based techniques of prenatal diagnosis such as amniocentesis, chorionic villus sampling (CVS) and percutaneous umbilical blood sampling (PUBS) are invasive and present significant risks of fetal loss. Non-invasive prenatal diagnosis utilizing fetal cells circulating in maternal peripheral blood during pregnancy has received much attention since it poses no risk to the fetus. Although several fetal cell types have been targeted, the search has focused on fetal nucleated red blood cells (fNRBC). However, because of very low concentration of fNRBC in the maternal blood, interference by nucleated red blood cells of maternal origin (mNRBC) and the failure to find broadly applicable identifiers that can differentiate fNRBC from mNRBC reliable detection and isolation of viable cells in amounts sufficient for clinical use remains a monumental challenge. Fetal NRBC differ from maternal NRBC in both nuclear morphology and hemoglobin molecular structure. Recently the principal investigator and his colleagues at MIT have developed biomedical light scattering spectroscopy (LSS) and showed its unique ability to optically probe nuclear morphology without damaging cells. Our group has further advanced LSS and developed the confocal absorption and scattering spectroscopic technique (CLASS) capable of monitoring subcellular organelles in small confocal volumes. The objective of this research program is twofold. (1) We will use the existing CLASS equipment developed in our laboratory to identify fNRBC native optical biomarkers. (2) Based on these biomarkers, we will develop a fast, simple and robust optical technique which combines CLASS microscopy and optical tweezers in the same instrument and which will enable enrichment and recovery of intact fNRBC from a sample of maternal peripheral blood. This will lead to a clinically useful method for prenatal genetic testing.