光谱流式细胞术登场

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据国外GEN网站介绍，一种光谱流式细胞术的新型技术开始登场了。尽管流式细胞术经过了数十年的发展，但是其检测时使用光电倍增管、分色镜、带通滤光片等技术一直延用至今。这种新型的光谱流式细胞术将这些技术抛弃了，改为棱镜或光栅来进行光谱分解（具体技术这篇报道中也没有详细说明，应该属于一种保密的专利性技术），并结合一种数据分析技术将光谱中的各种荧光重新区分开，估计每种荧光的强度，替代了原来复杂的补偿技术。
应该说读了这则新闻之后，令人心潮澎湃，不知道流式细胞仪厂商什么时候能够将这项技术加入到流式细胞仪中？拭目以待！

测试示意图

上图为利用La Jolla Bioengineering Institute’s 光谱流式细胞术做免疫分型的示例：外周血单个核细胞用荧光抗体标记，检测其全部发射光谱，通过参照光谱（即各种单色beads依次检测后合成的对照），可计算出细胞表面的荧光表达情况



【以下是新闻原稿】
Spectral Flow Cytometry Makes Debut

Although flow cytometry was invented more than 45 years ago, it continues to grow and expand into new areas that marry it with other modern cutting-edge technologies.

Harnessing the power of the full spectrum, coupling it with mass spectrometry, and utilizing it for expression profiling are a few of the new and intriguing applications heralding the next generation of flow cytometry.

Recent advances in optics and detectors are allowing a new technology, spectral flow cytometry, to make full spectral measurements on the sub-millisecond scale. John Nolan, Ph.D., principal investigator at La Jolla Bioengineering Institute, describes his approach: “There has been a longstanding interest in measuring the complete emission spectra from individual cells. We’ve developed the instrumentation to do this in a robust and routine way.”

Dr. Nolan says the new technology differs in several important aspects from conventional flow cytometry. “Instead of utilizing photomultiplier tubes, dichroic mirrors, and band-pass filters to measure cell-derived fluorescence at specific wavelengths, spectral flow cytometry uses prisms or gratings to disperse light over a detector array for high-speed, wavelength-resolved detection. Spectral unmixing, a data-analysis approach that estimates the amount of each fluorescent probe in the mixture spectrum, replaces compensation, which is required in conventional flow cytometry to account for fluorescence spillover between channels.”

• Spectral Flow
  Although autofluorescence can be a significant problem in typical flow cytometry, spectral flow easily resolves the issue. “With spectral flow, autofluorescence is just treated as another ‘color’ and can be resolved from other colors using spectral unmixing, effectively reducing a major source of background,” notes Dr. Nolan.
  Dr. Nolan and colleagues demonstrated proof of principle by using calibrated beads stained with six different quantum dots to demonstrate the analytical performance of the instrument. To evaluate performance in a typical immunophenotyping application, they analyzed peripheral blood mononuclear cells stained with canonical surface markers used to discriminate lymphocyte subsets.
  “Spectral flow was clearly able to resolve CD14+ monocytes, CD3+ T cells, and CD4+ and CD8+ subsets,” says Dr. Nolan. “This shows that spectral flow cytometry has a dynamic range suitable for such conventional applications.”
  Dr. Nolan and colleagues, aware that the emergence of spectral flow cytometry opens the doors to using many different types of fluorescent and other optical probes, are tackling other challenges for this evolving technology. “At present, commercial options are designed for the conventional flow cytometry paradigm of one color per detector,” remarks Dr. Nolan. “Within 5 to 10 years, we will likely see development of new probes, improved instrumentation, new analytical software, and new applications that take advantage of the spectral flow cytometry approach.”
  
  
  

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http://www.sony.net/SonyInfo/News/Press/201206/12-082E/

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