Article

Cytometry: Today’s technology and tomorrow’s horizons

ImmunoTechnology Section, VRC, NIAID, NIH, 40 Convent Dr., Room 5509, Bethesda, MD 20817, United States
Methods (Impact Factor: 3.22). 07/2012; 57(3):251–258. DOI: 10.1016/j.ymeth.2012.02.009

ABSTRACT Flow cytometry has been the premier tool for single cell analysis since its invention in the 1960s. It has maintained this position through steady advances in technology and applications, becoming the main force behind interrogating the complexities of the immune system. Technology development was a three-pronged effort, including the hardware, reagents, and analysis algorithms to allow measurement of as many as 20 independent parameters on each cell, at tens of thousands of cells per second. In the coming years, cytometry technology will integrate with other techniques, such as transcriptomics, metabolomics, and so forth. Ongoing efforts are aimed at algorithms to analyse these aggregated datasaets over large numbers of samples. Here we review the development efforts heralding the next stage of flow cytometry.

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    • "As mentioned above, flow cytometry has been the leading method for measuring this diversity over the past three decades. The latest instruments are remarkably powerful, capable of analyzing and/or sorting cells based on up to ∼18 cellular parameters at >10,000 cells per second4. Such high-dimensional and high-throughput analysis revealed relationships between T-cell phenotype/function and clinical status for a range of diseases. "
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    Nature Biotechnology 01/2014; 32(2). DOI:10.1038/nbt.2783 · 39.08 Impact Factor
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    • "Building cytometry panels (either user-assembled or with pre-prepared lyophilized reagent cocktails, such as Lyoplates™ from BD Biosciences) should include critical consideration of monoclonal antibody clones and the appropriate conjugate fluorescent detection dyes, as well as maintaining and monitoring consistent reagent performance as has been elaborated elsewhere (Kaminski et al., 2012; Maecker et al., 2012). Also described elsewhere are instrument set-up considerations and the extent of controls needed, which are familiar to many flow cytometry users (Perfetto et al., 2006; Maecker et al., 2010, 2012; Chattopadhyay and Roederer, 2012), but become extremely important to maintain consistency in large studies. These elements include single-color instrument compensation controls, fluorescence-minus-one controls that help determine the lower-limits of a positive stain when gating, and monitoring and documenting instrument set-up and performance on different procedure days of a study. "
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    Frontiers in Immunology 10/2012; 3:302. DOI:10.3389/fimmu.2012.00302
  • Methods 07/2012; 57(3):249-50. DOI:10.1016/j.ymeth.2012.08.010 · 3.22 Impact Factor
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