Microchip-based immunomagnetic detection of circulating tumor cells.
Microchip-based immunomagnetic detection of circulating tumor cells.
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DOI：
10.1039/c1lc20270g
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发表时间：
2011-10-21
期刊：
影响因子：
6.1
通讯作者：
Zhang X
中科院分区：
文献类型：
--
作者：
Hoshino K;Huang YY;Lane N;Huebschman M;Uhr JW;Frenkel EP;Zhang X
筛查血液中的循环肿瘤细胞（CTCs）一直是人们感兴趣的目标，以寻找疾病进展的证据、疾病活动状态、分子变化的克隆演变的识别以及可能的癌症早期诊断。我们描述了一种基于微芯片的免疫磁性CTC检测的新方法，其中结合了免疫磁性测定和微流控装置的优点。当血液样本流过紧靠阵列磁体上方的微通道时，用磁性纳米颗粒标记的癌细胞与血流分离并沉积在玻璃盖玻片的底壁上，这允许用荧光显微镜直接观察捕获的细胞。使用固定在玻璃盖玻片上的基于聚二甲基硅氧烷（PDMS）的微通道来筛选血液样品。微通道的薄而平坦的尺寸，与交替极性的阵列磁体附近的尖锐磁场梯度相结合，导致标记细胞的有效捕获。与市售的CellSearch™系统相比，需要更少（25%）的磁性颗粒来实现相当的捕获率，而筛选速度（在10 mL/小时的最佳血液流速下）比先前报道的基于微通道的测定快五倍以上。对于筛选实验，将从健康受试者抽取到CellSave™管中的血液掺入C 0 L0205和SKBR 3的培养的癌细胞系。然后在筛选前将血液在室温下保存48小时，模拟血液筛选的实际临床病例。将与抗上皮细胞粘附分子（EpCAM）抗体缀合的定制Fe 3 O 4磁性纳米颗粒（Veridex Ferrofluid™）引入血液样品中以标记癌细胞，然后使血液通过微芯片装置以捕获标记的细胞。捕获后，用荧光标记的抗细胞角蛋白、DAPI和抗CD 45对细胞进行染色。随后对捕获的细胞拍摄免疫荧光图像，然后基于荧光强度和细胞形态进行全面的计算机辅助分析。成功检测到罕见的癌细胞（从约1000个细胞降至约5个细胞/mL），肿瘤细胞与血细胞的比例非常低（约1：107~109，包括红细胞）。对COLO 205和SKBR 3细胞的癌细胞捕获率分别为90%和86%。
Screening for circulating tumor cells (CTCs) in blood has been an object of interest for evidence of progressive disease, status of disease activity, recognition of clonal evolution of molecular changes and for possible early diagnosis of cancer. We describe a new method of microchip-based immunomagnetic CTC detection, in which the benefits of both immunomagnetic assay and the microfluidic device are combined. As the blood sample flows through the microchannel closely above arrayed magnets, cancer cells labeled with magnetic nanoparticles are separated from blood flow and deposited at the bottom wall of the glass coverslip, which allows direct observation of captured cells with a fluorescence microscope. A polydimethylsiloxane (PDMS)-based microchannel fixed on a glass coverslip was used to screen blood samples. The thin, flat dimensions of the microchannel, combined with the sharp magnetic field gradient in the vicinity of arrayed magnets with alternate polarities, lead to an effective capture of labeled cells. Comparing to the commercially available CellSearch™ system, less (25%) magnetic particles are required to achieve a comparable capture rate, while the screening speed (at optimal blood flow rate of 10 mL/hour) is more than five times faster than those reported previously with a microchannel-based assay. For the screening experiment, blood drawn from healthy subjects into CellSave™ tubes was spiked with cultured cancer cell lines of COLO205 and SKBR3. The blood was then kept at room temperature for 48 hours before the screening, emulating the actual clinical cases of blood screening. Customized Fe3O4 magnetic nanoparticles (Veridex Ferrofluid™) conjugated to anti-Epithelial cell adhesion molecule (EpCAM) antibodies were introduced into the blood samples to label cancer cells, and the blood was then run through the microchip device to capture the labelled cells. After capture, the cells were stained with fluorescently labelled anti-cytokeratin, DAPI and anti-CD45. Subsequent immunofluorescence images were taken for the captured cells, followed by comprehensive computer aided analysis based on fluorescence intensities and cell morphology. Rare cancer cells (from ~1000 cells down to ~5 cells per mL) with very low tumor cell to blood cell ratios (about 1: 107~109, including red blood cells) were successfully detected. Cancer cell capture rates of 90% and 86% were demonstrated for COLO205 and SKBR3cells, respectively.
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影响因子：
6.4
作者：
He, Wei;Kularatne, Surnith A.;Kalli, Kimberly R.;Prendergast, Franklyn G.;Amato, Robert J.;Klee, George G.;Hartmann, Lynn C.;Low, Philip S.
通讯作者：
Low, Philip S.
影响因子：
3
作者：
Karl S;David M;Moore L;Grimberg BT;Michon P;Mueller I;Zborowski M;Zimmerman PA
通讯作者：
Zimmerman PA
影响因子：
2.8
作者：
Choi, Jin-Woo;Oh, Kwang W.;Ahn, Chong H.
通讯作者：
Ahn, Chong H.
DOI：
10.1158/1078-0432.ccr-10-1105
发表时间：
2010-10-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Lin HK;Zheng S;Williams AJ;Balic M;Groshen S;Scher HI;Fleisher M;Stadler W;Datar RH;Tai YC;Cote RJ
通讯作者：
Cote RJ
DOI：
10.1073/pnas.95.8.4589
发表时间：
1998-04-14
影响因子：
11.1
作者：
Racila, E;Euhus, D;Uhr, JW
通讯作者：
Uhr, JW