ImmunoTechnology Section, Vaccine Research Center, NIAID, NIH, Bethesda, Maryland 20892, USA.
Cytometry A. 2012 Jun;81(6):456-66. doi: 10.1002/cyto.a.22043. Epub 2012 Apr 6.
The Nobel Prize in Chemistry was awarded in 2000 for the discovery of conductive organic polymers, which have subsequently been adapted for applications in ultrasensitive biological detection. Here, we report the first use of this new class of fluorescent probes in a diverse range of cytometric and imaging applications. We demonstrate that these "Brilliant Violet" reporters are dramatically brighter than other UV-violet excitable dyes, and are of similar utility to phycoerythrin (PE) and allophycocyanin (APC). They are thus ideally suited for cytometric assays requiring high sensitivity, such as MHC-multimer staining or detection of intracellular antigens. Furthermore, these reporters are sensitive and spectrally distinct options for fluorescence imaging, two-photon microscopy and imaging cytometry. These ultra-bright materials provide the first new high-sensitivity fluorescence probes in over 25 years and will have a dramatic impact on the design and implementation of multicolor panels for high-sensitivity immunofluorescence assays.
2000 年诺贝尔化学奖授予对导电有机聚合物的发现,随后这些聚合物被应用于超灵敏生物检测。在这里,我们报告了这一新类荧光探针在各种细胞测量和成像应用中的首次使用。我们证明,这些“亮艳紫”报告分子比其他紫外-可见激发染料亮得多,并且与藻红蛋白(PE)和别藻蓝蛋白(APC)的用途相似。因此,它们非常适合需要高灵敏度的细胞测量应用,例如 MHC 多聚体染色或细胞内抗原的检测。此外,这些报告分子是荧光成像、双光子显微镜和成像细胞测量的敏感且光谱独特的选择。这些超高亮度材料提供了 25 年来第一批新的高灵敏度荧光探针,将对高灵敏度免疫荧光测定中多色面板的设计和实施产生巨大影响。