使用量子点通过扫描荧光显微镜对白细胞进行自动四色分析。
Automated four-color analysis of leukocytes by scanning fluorescence microscopy using quantum dots.
作者信息
Bocsi József, Lenz Dominik, Mittag Anja, Varga Viktor Sebestyén, Molnar Béla, Tulassay Zsolt, Sack Ulrich, Tárnok Attila
机构信息
Department of Pediatric Cardiology, Heart Center, University Leipzig, Leipzig, Germany.
出版信息
Cytometry A. 2006 Mar;69(3):131-4. doi: 10.1002/cyto.a.20217.
BACKGROUND
Scanning fluorescence microscope (SFM) is a new technique for automated motorized microscopes to measure multiple fluorochrome labeled cells (Bocsi et al., Cytometry A 2004, 61:1-8).
AIMS
We developed a four-color staining protocol (DNA, CD3, CD4, and CD8) for the lymphocyte phenotyping by SFM.
METHODS
Organic (Alexa488, FITC, PE-Alexa610, CyChrom, APC) and inorganic (quantum dot (QD) 605 or 655) fluorochromes were used and compared in different combinations. Measurements were performed in suspension by flow cytometer (FCM) and on slide by SFM.
RESULTS
Both QDs were detectable by the appropriate Axioplan-2 and FCM filters and the AxioCam BW-camera. CD4/CD8 ratios were highly correlated (P = 0.01) between the SFM and FCM.
CONCLUSION
Automated SFM is an applicable tool for CD4/CD8 ratio determination in peripheral blood samples with QDs.
背景
扫描荧光显微镜（SFM）是自动电动显微镜用于测量多种荧光染料标记细胞的一项新技术（博奇等人，《细胞分析A》2004年，61:1 - 8）。
目的
我们开发了一种用于通过SFM进行淋巴细胞表型分析的四色染色方案（DNA、CD3、CD4和CD8）。
方法
使用有机（Alexa488、FITC、PE - Alexa610、CyChrom、APC）和无机（量子点（QD）605或655）荧光染料，并对不同组合进行比较。通过流式细胞仪（FCM）在悬浮液中以及通过SFM在载玻片上进行测量。
结果
两种量子点均可通过合适的Axioplan - 2和FCM滤光片以及AxioCam BW相机检测到。SFM和FCM之间的CD4/CD8比值高度相关（P = 0.01）。
结论
自动化SFM是一种适用于使用量子点测定外周血样本中CD4/CD8比值的工具。
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