Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function.
Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function.
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DOI：
10.1021/ac100142a
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发表时间：
2010-05-01
影响因子：
7.4
通讯作者：
Revzin, Alexander
中科院分区：
文献类型：
--
作者：
Stybayeva, Gulnaz;Mudanyali, Onur;Seo, Sungkyu;Silangcruz, Jaime;Macal, Monica;Ramanculov, Erlan;Dandekar, Satya;Erlinger, Anthony;Ozcan, Aydogan;Revzin, Alexander
白细胞的特征是血液分析和基于血液的诊断的一个组成部分。在本文中，我们将无透镜全息成像与抗体微阵列相结合，用于快速和多参数分析人血液中的白细胞。将白细胞表面抗原（CD4和CD8）和细胞因子（TNF-α、IFN-γ、IL-2）特异性单克隆抗体（Abs）打印成阵列，并置细胞捕获和细胞因子检测Ab点。将Ab微阵列整合到4 μl体积的微流控室中，然后与人血液孵育，在特定的Ab点上捕获CD4和CD8 t细胞。芯片上这些细胞的有丝分裂激活诱导细胞因子分子的释放，这些分子随后被捕获到邻近的抗细胞因子Ab点上。使用hrp标记的抗细胞因子抗体和可见色试剂检测IL-2、TNF-α和IFN-γ分子在各自Ab点上的结合。然后使用无透镜全息成像快速（~ 4秒）枚举Ab点上捕获的CD4和CD8 t淋巴细胞，并量化同一芯片上IL-2、TNF-α和IFN-γ点发出的细胞因子信号。为了证明我们的方法在传染病监测中的实用性，我们分析了健康志愿者和人类免疫缺陷病毒（HIV）感染患者的血液样本，以确定CD4/CD8比率——一个重要的HIV/AIDS诊断指标。通过在Ab点上捕获的CD4和CD8 t细胞的无透镜芯片成像获得的比例与传统的基于显微镜的细胞计数非常一致。本论文描述了Ab微阵列和无透镜全息成像的串联使用，为未来在护理点或资源有限的环境中用于多参数血液分析的微型细胞仪设备的发展铺平了道路。
Characterization of leukocytes is an integral part of blood analysis and blood-based diagnostics. In the present paper we combine lensless holographic imaging with antibody microarrays for rapid and multiparametric analysis of leukocytes from human blood. Monoclonal antibodies (Abs) specific for leukocyte surface antigens (CD4 and CD8) and cytokines (TNF-α, IFN-γ, IL-2) were printed in an array so as to juxtapose cell capture and cytokine detection Ab spots. Integration of Ab microarrays into a microfluidic flow chamber (4 μl volume) followed by incubation with human blood resulted in capture of CD4 and CD8 T-cells on specific Ab spots. On-chip mitogenic activation of these cells induced release of cytokine molecules that were subsequently captured on neighboring anti-cytokine Ab spots. The binding of IL-2, TNF-α and IFN-γ molecules on their respective Ab spots was detected using HRP-labeled anti-cytokine Abs and a visible color reagent. Lensfree holographic imaging was then used to rapidly (∼4 sec) enumerate CD4 and CD8 T-lymphocytes captured on Ab spots and to quantify the cytokine signal emanating from IL-2, TNF-α, and IFN-γ spots on the same chip. To demonstrate the utility of our approach for infectious disease monitoring, blood samples of healthy volunteers and human immunodeficiency virus (HIV)-infected patients were analyzed to determine CD4/CD8 ratio – an important HIV/AIDS diagnostic marker. The ratio obtained by lensfree on-chip imaging of CD4 and CD8 T-cells captured on Ab spots was in close agreement with conventional microscopy-based cell counting. The present paper, describing tandem use of Ab microarrays and lensfree holographic imaging, paves the way for future development of miniature cytometry devices for multiparametric blood analysis at the point of care or in a resource-limited setting.
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影响因子：
6.1
作者：
Seo S;Su TW;Tseng DK;Erlinger A;Ozcan A
通讯作者：
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影响因子：
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影响因子：
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通讯作者：
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影响因子：
2.8
作者：
Seo, Sungkyu;Su, Ting-Wei;Ozcan, Aydogan
通讯作者：
Ozcan, Aydogan
影响因子：
15.8
作者：
Rodriguez, WR;Christodoulides, N;Floriano, PN;Graham, S;Mohanty, S;Dixon, M;Hsiang, M;Peter, T;Zavahir, S;Thior, I;Romanovicz, D;Bernard, B;Goodey, AP;Walker, BD;McDevitt, JT
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McDevitt, JT