Nanosensor Chemical Cytometry for Characterizing the Efflux Heterogeneity of Nitric Oxide from Macrophages
Nanosensor Chemical Cytometry for Characterizing the Efflux Heterogeneity of Nitric Oxide from Macrophages
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DOI：
10.1021/acsnano.1c04958
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发表时间：
2021-08-16
期刊：
影响因子：
17.1
通讯作者：
Strano，Michael S.
中科院分区：
文献类型：
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作者：
Cho，Soo-Yeon;Koman，Volodymyr B.;Strano，Michael S.
巨噬细胞是人类免疫反应的关键部分，它们的集体异质性与疾病的进展和预防有关。尽管有诊断和治疗的潜力，但目前还不存在非破坏性、无标记的工具来描述集合中单个巨噬细胞的动态抗原反应，以与细胞水平上特定的分子表达和相关的生物物理性质相关联。在这里，我们发展了一种纳米传感器化学细胞术(NCC)，可以描述巨噬细胞群体诱导型一氧化氮合酶(INOS)反应的异质性。通过将近红外(NIR)荧光纳米传感器阵列和胶原层与微流体相结合，利用巨噬细胞的细胞透镜效应在单细胞水平上表征一氧化氮(NO)外流和折射率(RI)的变化。使用并行的多通道方法，可以在大摩尔(10-18mol)灵敏度下以无损和实时的方式监测巨噬细胞不同的iNOS异质性，吞吐量超过200个细胞/帧。我们发现，巨噬细胞群体的估计平均NO外流率从342(σ=199)增加到464(σ=206)attomol/cell·hr，异质性增加了3%，并且巨噬细胞的估计RI从1.366(σ=0.015)下降到1.359(σ=0.009)，其中三种亚群的巨噬细胞在内毒素激活下。这些测量值也与Griess化验结果和先前报道的测量结果很好地一致。这项工作为细胞制造和生物制药工程中的巨噬细胞群体的单细胞分析提供了一种有效的策略。
Macrophages are a critical part of the human immune response, and their collective heterogeneity is implicated in disease progression and prevention. A nondestructive, label-free tool does not currently exist for profiling the dynamic, antigenic responses of single macrophages in a collection to correlate with specific molecular expression and correlated biophysical properties at the cellular level, despite the potential for diagnosis and therapeutics. Herein, we develop a nanosensor chemical cytometry (NCC) that can profile the heterogeneity of inducible nitric oxide synthase (iNOS) responses from macrophage populations. By integrating a near-infrared (nIR) fluorescent nanosensor array and collagen layer with microfluidics, the cellular lensing effect of the macrophage was utilized to characterize both nitric oxide (NO) efflux and refractive index (RI) changes at a single-cell level. Using a parallel, multichannel approach, distinct iNOS heterogeneities of macrophages can be monitored at an attomolar (10–18mol) sensitivity in a nondestructive and real-time manner with a throughput of exceeding the 200 cells/frame. We demonstrate that estimated mean NO efflux rates of macrophage populations are elevated from 342 (σ = 199) to 464 (σ = 206) attomol/cell·hr with a 3% larger increase in the heterogeneity, and estimated RI of macrophage decrease from 1.366 (σ = 0.015) to 1.359 (σ = 0.009) with trimodal subpopulations under lipopolysaccharide (LPS) activation. These measured values are also in good agreement with Griess assay results and previously reported measurements. This work provides an efficient strategy for single-cell analysis of macrophage populations for cellular manufacturing and biopharmaceutical engineering.