Molecular Imaging Program at Stanford (MIPS), Department of Radiology, Stanford University, Stanford, CA, 94305, USA.
Adv Biol (Weinh). 2021 Mar;5(3):e2000441. doi: 10.1002/adbi.202000441. Epub 2021 Jan 27.
Here, a method for label-free, real-time interrogation, monitoring, detection, and sorting of biological rare cells in magnetically suspended heterogeneous samples is developed. To achieve this, heterogeneous populations of cells are levitated and confined in a microcapillary channel. This strategy enables spatiotemporal differential magnetic levitation of rare fragile dead cells equilibrating at different heights based on the balance between magnetic and corrected gravitational forces. In addition, the sorting of fragile rare dead cell populations is monitored in real-time. This technique provides a broadly applicable label-free tool for high resolution, real-time research, as well as forensic evidence processing of rape kits. This method is validated with forensic mock samples dating back to 2003, isolating sperm from epithelial cells (E. cells) with >90% efficiency and >97% purity. Overall, this method reduces the processing time by over 20-fold down to 20 min, eliminating centrifugation and labels, and providing an inexpensive and high-yield alternative to the current centrifuge-based differential extraction techniques. It can potentially facilitate the forensic downstream genomic analyses, accelerating the identification of suspects, and advancing public safety.
本文开发了一种无标记、实时询问、监测、检测和分选悬浮在异质样品中的生物稀有细胞的方法。为了实现这一目标，将异质细胞群体悬浮并限制在微流控通道中。这种策略能够实现稀有易碎死细胞的时空差分磁悬浮，根据磁场和修正重力之间的平衡，使它们在不同的高度达到平衡。此外，还可以实时监测易碎稀有死细胞群体的分选。该技术为高分辨率、实时研究以及强奸工具包的法医证据处理提供了一种广泛适用的无标记工具。该方法用可追溯到 2003 年的法医模拟样本进行了验证，从上皮细胞（E.细胞）中分离精子的效率超过 90%，纯度超过 97%。总的来说，这种方法将处理时间缩短了 20 多倍，达到 20 分钟，消除了离心和标记，并为当前基于离心的差异提取技术提供了一种廉价且高产的替代方法。它有可能促进法医下游的基因组分析，加速嫌疑人的识别，提高公共安全。
