Boribong Brittany P, Rahimi Amina, Jones Caroline N
Genetics, Bioinformatics, and Computational Biology, Virginia Tech, Blacksburg, VA, USA.
Department of Biochemistry, Virginia Tech, Blacksburg, VA, USA.
Methods Mol Biol. 2019;1960:113-122. doi: 10.1007/978-1-4939-9167-9_10.
Neutrophils are the most abundant leukocytes in blood, serving as the first line of host defense in tissue damage and infections. Upon activation by chemokines released from pathogens or injured tissues, neutrophils migrate through complex tissue microenvironments toward sites of infections along the chemokine gradients, in a process named chemotaxis. However, current methods for measuring neutrophil chemotaxis require large volumes of blood and are often bulk, endpoint measurements. To address the need for rapid and robust assays, we engineered a novel dual gradient microfluidic platform that precisely quantifies neutrophil migratory decision-making with high temporal resolution. Here, we present a protocol to measure neutrophil migratory phenotypes (velocity, directionality) with single-cell resolution.
中性粒细胞是血液中数量最多的白细胞，在组织损伤和感染中作为宿主防御的第一道防线。在病原体或受损组织释放的趋化因子激活后，中性粒细胞沿着趋化因子梯度穿过复杂的组织微环境，向感染部位迁移，这一过程称为趋化作用。然而，目前测量中性粒细胞趋化作用的方法需要大量血液，而且往往是整体的终点测量。为了满足对快速且可靠检测方法的需求，我们设计了一种新型双梯度微流控平台，该平台能够以高时间分辨率精确量化中性粒细胞的迁移决策。在此，我们展示一种以单细胞分辨率测量中性粒细胞迁移表型（速度、方向性）的方案。
