Development and use of nanoFACS for analysis and sorting of nanoparticles

我们成功地将nanoFACS（一种用于分析和分选naonoscale颗粒的方法）引入NIH，通过修改10号楼NCI/CCR核心FACS设施中的Influx流式细胞仪。现在，这使我们能够分析和分选小至约100 nm的外来体和微粒。我们已经开发并测试了技术和仪器配置的许多修改，以提高检测限，以常规分析和分选小至40 nm的颗粒（这将涵盖生物学上重要的外泌体尺寸的全部范围）。我们还获得了一台新仪器，Beckman Coutler MoFlo AstriosEQ，经过修改，进一步提高了我们的分析和分选能力，作为我们集团与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型生物标志物，我们可以用它们来指导和监测癌症治疗和免疫反应（对疫苗或其他免疫疗法）的影响，和2）识别这些纳米颗粒的子集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.