Multiplex cytometry

McDade R.L., Fulton R.J., True multiplexed analysis by computer-enhanced flow cytometry, Medical Device and Diagnostic Industry, 6 pp., April 1977. 

Armstrong, B. et al. (2000) Suspension arrays for high throughput, multiplexed single nucleotide polymorphism genotyping. Cytometry 40, 102-108. 

Lowe M, Spiro A, Zhang YZ et al (2004) Multiplexed, particle-based detection of DNA using flow cytometry with 3DNA dendrimers for signal amplification. Cytometry Part A 60A 135-144... 

Nolan J, Mandy F (2006) Multiplexed and microparticle-based analyses. Quantitative tools for the large-scale analysis of biological systems. Cytometry A 69 318-325... 

Flow cytometry Allows the quantification of the proportion of cells expressing a certain antigen/s Multiplexing capability. Cells must be detached from material prior to analysis Yes... 

Results using multiplexed bead flow cytometry have been reported by Carson RT, Vignali DAA (1999). Simultaneous quantitation of 15 cytokines using a multiplexed flow cytometric assay. J. Immunol. Methods 227 41-52. 

Iannone MA, Taylor JD, Chen J, Li MS, Rivers P, Slentz-Kesler KA, Weiner MP. Multiplexed single nucleotide polymorphism genotyping by oligonucleotide ligation and flow cytometry. Cytometry 2000 39 131-140. 

Luminex is also a bead-based, non-separation technology using the Luminex colour-coded beads and detection systems (Luminex 100 IS or Luminex HT ). The readers used for this assay format are based on the principle of flow cytometry. The system enables assays to be multiplexed, i.e. allowing different analytes to be monitored simultaneously. The Luminex HT system is compatible with 96- and 384-well microplates14 but throughput of the reader is still a limiting factor for large-scale HTS. 

Interesting studies were also performed based on a suspension of beads in conjunction with flow cytometry measurements [23,24], Flow cytometry, which was the standard methodology for cell population study during the last 20 years, has now begun to serve for in vitro microspheres analysis [25]. Such systems were described as multiplex microsphere bead assays and were used to detect different nucleic acid sequences hybridized on beads having different properties (size, fluorescent label). 

Pataki J, Szabo M, Lantos E, Szekvolgyi L, Molnar M, Hegedus E, Bacso Z, Kappelmayer J, Lustyik G, Szabo G. Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR. Cytometry Part A 2005 68A 45-52. Martins TB, Augustine NH, Hill HR. Development of a multiplexed fluorescent immunoassay for the quantitation of antibody responses to group A streptococci. J. Immunol. Methods 2006 316 97-106. 

Cell-Based Bioassay For a cell-based Nab bioassay, treated cells respond directly or indirectly to the drug in a concentration-dependent manner. Possible biological responses of the cells to chug treatment include cell proliferation, apoptosis, phosphorylation, chemokine release, and expression of proteins or genes [15,22,28 30]. These responses may by quantitated by techniques such as immunoassay, multiplex assays [31], flow cytometry [23], and gene expression profiling [32]. 

KeUar KL, Kalwar RR, Dubois KA, Crouse D, Chafin WD, Kane BE. Multiplexed fluorescent bead-based immunoassays for quantitation of human cytokines in serum and culture supernatants. Cytometry 2001 45 27-36. 

In microbead arrays, capture molecules attached onto microbead surfaces of, for example, polystyrene microspheres, act as protein microarrays and thus, can be used as such in protein-protein, nucleic acid-protein, and nucleic acid-nucleic acid interaction studies using flow cytometry-based interrogation. Multiplexing is possible by the utilization of sized [20] or color-coded microbeads [155]. 

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