...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Lab on a chip >Droplet encapsulation improves accuracy of immune cell cytokine capture assays
【24h】

Droplet encapsulation improves accuracy of immune cell cytokine capture assays

机译：液滴封装改善了免疫细胞细胞因子捕获测定的准确性

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Quantification of cell-secreted molecules, e.g., cytokines, is fundamental to the characterization of immune responses. Cytokine capture assays that use engineered antibodies to anchor the secreted molecules to the secreting cells are widely used to characterize immune responses because they allow both sensitive identification and recovery of viable responding cells. However, if the cytokines diffuse away from the secreting cells, non-secreting cells will also be identified as responding cells. Here we encapsulate immune cells in microfluidic droplets and perform in-droplet cytokine capture assays to limit the diffusion of the secreted cytokines. We use microfluidic devices to rapidly encapsulate single natural killer NK-92 MI cells and their target K562 cells into microfluidic droplets. We perform in-droplet IFN-gamma capture assays and demonstrate that NK-92 MI cells recognize target cells within droplets and become activated to secrete IFN-gamma. Droplet encapsulation prevents diffusion of secreted products to neighboring cells and dramatically reduces both false positives and false negatives, relative to assays performed without droplets. In a sample containing 1% true positives, encapsulation reduces, from 94% to 2%, the number of true-positive cells appearing as negatives; in a sample containing 50% true positives, the number of non-stimulated cells appearing as positives is reduced from 98% to 1%. After cells are released from the droplets, secreted cytokine remains captured onto secreting immune cells, enabling FACS-isolation of populations highly enriched for activated effector immune cells. Droplet encapsulation can be used to reduce background and improve detection of any single-cell secretion assay.
机译：细胞分泌分子的定量，例如细胞因子是对免疫反应表征的基础。使用工程化抗体将分泌物分泌到分泌细胞的细胞因子捕获测定被广泛用于表征免疫应答，因为它们允许敏感的鉴定和恢复可行的响应细胞。然而，如果细胞因子扩散远离分泌细胞，则也将鉴定非分泌细胞作为响应细胞。在这里，我们将免疫细胞封装在微流体液滴中并进行液滴细胞因子捕获测定以限制分泌细胞因子的扩散。我们使用微流体装置将单一天然杀伤NK-92 MI细胞及其靶K562细胞迅速封装到微流体液滴中。我们在液滴IFN-Gamma捕获测定中表明NK-92 MI细胞识别液滴内的靶细胞，并被激活分泌IFN-γ。液滴封装可防止分泌产物的扩散到相邻细胞中，并且相对于没有液滴的测定，显着降低了误报和假阴性。在含有1％真实阳性的样品中，封装减少，从94％到2％，出现为底片的真阳性细胞的数量;在含有50％真实阳性的样品中，出现为阳性的非刺激细胞的数量从98％降至1％。在从液滴中释放细胞后，分泌的细胞因子仍然捕获到分泌物免疫细胞上，使得能够对活性效应剂免疫细胞高度富集的群体分离。液滴封装可用于减少背景和改善任何单细胞分泌测定的检测。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号