Development and use of nanoFACS for analysis and sorting of nanoparticles

我们通过对位于10号楼的NCI/CCR核心FACS设施中的流入流式细胞仪进行改进，成功地将纳米FACS引入NIH，这是一种分析和分选纳米尺度颗粒的方法。现在，我们可以分析和分选小至约100 nm的外切体和微粒。我们已经开发和测试了对技术和仪器配置的大量修改，以提高检测极限，以便对小至40 nm的颗粒进行常规分析和分类(这将涵盖所有具有生物学意义的外体大小)。我们还获得了一台新仪器Beckman Coutler MoFlo AstriosEQ，经过改进以进一步提高我们的分析和分类能力，这是我们集团与Beckman之间建立的CRADA的一部分。在接下来的一年里，我们将继续1)使用这项技术来识别新的生物标记物，我们可以使用这些生物标记物来指导和监测癌症治疗的效果和(对疫苗或其他免疫疗法的)免疫反应，以及2)识别这些纳米颗粒的亚群，并确定它们在肿瘤生物学、免疫生物学和生物学其他方面的功能。

We successfully brought nanoFACS, a method for analyzing and sorting naonoscale particles, to NIH, by modifying the Influx flow cytometer in a NCI/CCR core FACS facility in Building 10. This now allows us to analyze and sort exosomes and microparticles as small as approximately 100nm. We have developed and tested numerous modifications of the technique and instrument configuration to improve the detection limits to routinely analyze and sort particles as small as 40nm (which would span the full range of biologically significant exosome sizes). We also have obtained a new instrument, a Beckman Coutler MoFlo AstriosEQ,with modifications to further improve our analystical and sorting capabilities, as part of a CRADA established between our group and Beckman. Over the next year, we will continue 1) to use this technology to identify novel biomarkers that we can use to guide and monitor the effects of cancer therapies and immune responses (to vaccines or other immunotherapies), and 2) to identify subsets of these nanoparticles and determine their functions in tumor biology, immunobiology, and other aspects of biology.