High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles-Brief Report
High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles-Brief Report
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DOI：
10.1161/atvbaha.111.244616
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发表时间：
2012-04-01
影响因子：
8.7
通讯作者：
Dignat-George, Francoise
中科院分区：
文献类型：
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作者：
Robert, Stephane;Lacroix, Romaric;Dignat-George, Francoise
微细胞微粒（MP）是许多病理情况下有前途的生物标志物。虽然流式细胞术（FCM）被广泛用于它们的测量，但它引起了争议，因为最小的MP大小福尔斯标准FCM（sd-FCM）的检测限。在最近的技术改进导致高灵敏度FCM（hs-FCM）之后，我们的目标是（1）评估hs-FCM用于扩展MP检测的潜力，（2）建立MP计数的标准化方案，和（3）比较用两种灵敏度水平获得的MP计数。使我们能够区分血浆样本中以前无法检测到的小MP。使用具有适当大小（0.1/0.3/0.5/0.9 μ m）和相对量的荧光珠，建立了新的标准化hs-FCM MP方案，并提供了可重复的MP计数。应用于冠状动脉患者样本，与sd-FCM相比，它导致MP计数增加8至20倍。有趣的是，小和大MP之间的比例根据临床状态而变化，但也取决于MP子集，这表明获得新的生物信息。结论-FCM的最新改进提供了以前无法检测到的MP，并代表了一个新的机会，以提高其作为生物标志物在临床实践中的影响。（Arterioscler Thromb Vasc Biol.2012;32：1054-1058.）
Objective-Cellular microparticles (MP) are promising biomarkers in many pathological situations. Although flow cytometry (FCM) is widely used for their measurement, it has raised controversies because the smallest MP size falls below the detection limit of standard FCM (sd-FCM). Following recent technological improvements leading to high sensitivity FCM (hs-FCM), our objectives were (1) to evaluate the potential of hs-FCM for extended MP detection, (2) to set up a standardized protocol for MP enumeration, and (3) to compare MP counts obtained with both sensitivity levels.Methods and Results-Compared with sd-FCM, hs-FCM displayed improved forward scatter resolution and lower background noise, allowing us to discriminate previously undetectable small MP in plasma samples. Using fluorescent beads with appropriate sizes (0.1/0.3/0.5/0.9 mu m) and relative amounts, a new standardized hs-FCM MP protocol was set up and provided reproducible MP counts. Applied to coronary patient samples, it resulted into 8- to 20-fold increases in MP counts as compared with sd-FCM. Interestingly, the ratio between small and large MP varied according to clinical status but also depending on MP subset, suggesting access to new biological information.Conclusion-Recent improvements in FCM provide access to previously undetectable MP and represent a new opportunity to enhance their impact as biomarkers in clinical practice. (Arterioscler Thromb Vasc Biol. 2012;32:1054-1058.)