Scanning cytometry with a LEAP: laser-enabled analysis and processing of live cells in situ.
Scanning cytometry with a LEAP: laser-enabled analysis and processing of live cells in situ.
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使用 LEAP 扫描细胞术：利用激光对活细胞进行原位分析和处理。
DOI：
10.1002/cyto.a.20291
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Leary,JamesF
中科院分区：
文献类型：
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作者：
Szaniszlo,Peter;Rose,WilliamA;Wang,Nan;Reece,LisaM;Tsulaia,TamaraV;Hanania,ElieG;Elferink,CornelisJ;Leary,JamesF
背景扫描式流式细胞术具有多参数流式细胞术的许多功能，同时又保持了自身作为一种成像技术的优势。现代仪器结合了联合收割机的扫描细胞计数能力和操作细胞的能力。一种新的技术，称为LEAP™（激光使能分析和处理），提供了一个独特的组合的能力，在细胞纯化和选择性大分子传递（optinjection）。MethodsLEAP介导的细胞纯化和optinjection的影响进行了评估，在模型实验中使用贴壁和悬浮细胞类型和细胞混合物铺板和处理在不同的密度。通过将荧光葡聚糖递送到细胞中来可视化光注射效应。结果进行了分析，使用LEAP仪器的自己的成像系统，以及通过荧光和共聚焦microscopic.ResultsLive细胞样品（贴壁和悬浮液）可以纯化到90-100%的纯度与50-90%的产量，造成最小的细胞损伤取决于细胞类型和平板密度。几乎100%的被检测细胞类型的靶细胞可以成功地用3-70 kDa的葡聚糖进行光注射，对细胞没有造成视觉损伤。间接optoinjection效果被观察到在5-60 μm内的非靶向细胞的目标areas under conditions used here.ConclusionsLEAP提供了解决方案，在细胞纯化和有针对性的大分子传递的传统和具有挑战性的应用程序，其他方法不足。© 2006国际分析细胞学学会
BackgroundScanning cytometry now has many of the features (and power) of multiparameter flow cytometry while keeping its own advantages as an imaging technology. Modern instruments combine capabilities of scanning cytometry with the ability to manipulate cells. A new technology, called LEAP™ (laser‐enabled analysis and processing), offers a unique combination of capabilities in cell purification and selective macromolecule delivery (optoinjection).MethodsLEAP‐mediated cell purification and optoinjection effects were assessed in model experiments using adherent and suspension cell types and cell mixtures plated and processed at different densities. Optoinjection effects were visualized by delivering fluorescent dextrans into cells. Results were analyzed using the LEAP instrument's own imaging system as well as by fluorescence and confocal microscopy.ResultsLive cell samples (adherent and suspension) could be purified to 90–100% purity with 50–90% yield, causing minimal cell damage depending on the cell type and plating density. Nearly one hundred percent of the targeted cells of all cell types examined could be successfully optoinjected with dextrans of 3–70 kDa, causing no visual damage to the cells. Indirect optoinjection effects were observed on untargeted cells within 5–60 μm to targeted areas under conditions used here.ConclusionsLEAP provides solutions in cell purification and targeted macromolecule delivery for traditional and challenging applications where other methods fall short. © 2006 International Society for Analytical Cytology