Quantitative measurement of multifunctional quantum dot binding to cellular targets using flow cytometry.
Quantitative measurement of multifunctional quantum dot binding to cellular targets using flow cytometry.
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使用流式细胞术对多功能量子点结合的多功能量子点结合。
DOI：
10.1002/cyto.a.20677
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发表时间：
2009-05
期刊：
影响因子：
--
通讯作者：
Giorgio TD
中科院分区：
文献类型：
--
作者：
Smith RA;Giorgio TD
量子点等半导体纳米晶在流式细胞仪和荧光显微镜领域是一种潜在的强大资源。量子点的大小和荧光特性为靶向递送系统和流式细胞仪检测提供了诱人的特征。虽然各种荧光分子的定量测量是常规的，但没有校准标准的荧光团，如量子点，给流式细胞仪的定量带来了问题。我们的目标是开发一种有针对性的纳米颗粒传递平台以及相应的方法来准确和定量地评估该系统的性能。我们合成了针对细胞表面受体的表面修饰的QD探针，并用流式细胞仪测量了所得到的细胞-探针偶联物的MFI。使用标准校准微球将MFI转换为平均等效R-PE强度(MEPE)。用荧光法测量R-PE和QD探针的已知浓度，以关联R-PE和QD荧光。然后使用荧光测量结果将MEPE测量结果转换为结合的QD探针的数量。靶向探针表现出比未修饰和非靶向颗粒更好的结合特性。这种结合作用被证明是特异的，并由连接到QD表面的NGR靶向多肽介导。为评估该系统而开发的校准方法被证明成功地将原始荧光数据转换为定量探针结合值。我们展示了一种高度模块化的纳米粒子系统的合成和性能，该系统能够进行靶向结合和荧光成像。为量化该系统的性能而实施的校准方法代表了一种潜在的强大工具，可以利用包括量子点在内的一系列荧光分子进行真正定量的流式细胞仪测量。
Semiconductor nanocrystals such as quantum dots (QDs) are a potentially powerful resource in the fields of flow cytometry and fluorescence microscopy. QD size and fluorescence characteristics offer attractive features for use in targeted delivery systems and detection by flow cytometry. While quantitative measurements of a variety of fluorescent molecules are routinely performed, fluorophores for which no calibration standards exist, such as QDs, pose a problem for quantitation in flow cytometry. Our goal was to develop a targeted nanoparticle delivery platform as well as a corresponding method to accurately and quantitatively assess the performance of this system. We synthesized surface-modified QD probes targeted to cellular surface receptors and measured the MFI of the resulting cell-probe conjugates by flow cytometry. MFI was converted to mean equivalent R-PE intensity (MEPE) using standard calibration microspheres. Known concentrations of both R-PE and QD probes were measured by fluorometry to relate R-PE and QD fluorescence. Fluorometry results were then used to translate MEPE measurements to the number of bound QD probes. The targeted probes exhibited superior binding characteristics over unmodified and untargeted particles. This binding interaction was shown to be specific and mediated by the NGR targeting peptide tethered to the QD surface. The calibration method developed to assess this system proved successful at converting raw fluorescence data to quantitative probe binding values. We demonstrate the synthesis and performance of a highly modular nanoparticle system capable of targeted binding and fluorescent imaging. The calibration method implemented to quantify the performance of this system represents a potentially powerful tool to utilize truly quantitative flow cytometry measurements with an array of fluorescent molecules, including QDs.
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DOI：
10.1073/pnas.152463399
发表时间：
2002-10-01
影响因子：
11.1
作者：
Åkerman, ME;Chan, WCW;Ruoslahti, E
通讯作者：
Ruoslahti, E
DOI：
10.1002/cyto.990220311
发表时间：
1995-09-15
期刊：
CYTOMETRY
影响因子：
--
作者：
CHANCE, JT;LARSEN, SA;COX, DL
通讯作者：
COX, DL
影响因子：
2.9
作者：
FAY, SP;POSNER, RG;SKLAR, LA
通讯作者：
SKLAR, LA
影响因子：
50.3
作者：
Hallahan, D;Geng, L;Clanton, J
通讯作者：
Clanton, J
影响因子：
46.9
作者：
Jaiswal, JK;Mattoussi, H;Simon, SM
通讯作者：
Simon, SM