Abstract
BackgroundFlow cytometry based suspended microarray assays are susceptible to many sources of variance; multi-well replication and inter-instrument reproducibility is uncertain.Method and resultsAn "intraplex" method was developed in order to minimize differences in sample readings between instruments. A full intraplex assay consists of a set m of microparticle set classifications assaying for the same analyte, with each of the m classifier sets having different sensitivity to analyte, and n classifier sets replicating each of the m levels of sensitivity, where m > 1 (generally m > 4 would be used).ConclusionThe intraplex method can compensate adequately for the sources of variance that have been identified in suspended microarray assays. It requires no changes to current equipment in use, and is a superior method of constructing precision assays. Additionally, Luminex® users may want to consider the evidence that shows that despite calibration to the same standard, two instruments may not give similar results for all concentrations of analytes.
Highlights
Flow cytometry based suspended microarray assays are susceptible to many sources of variance; multi-well replication and inter-instrument reproducibility is uncertain
Assay on the surface of a small particle.) The particles are run through a flow cytometer, which may be optimized for the specific assay system
The virus antigens used in these experiments were: CMV- Cytomegalovirus, SFV- Simian Foamy Virus, The full intraplex conceptualized in Figure 2 is composed of an m × n matrix in which each of m different SMPCSIDG's has n Suspended Microarray Particle Classifier Set (SMPCS)'s designed to be identical
Summary
Flow cytometry based suspended microarray assays are susceptible to many sources of variance; multi-well replication and inter-instrument reproducibility is uncertain. A suspended microarray assay system uses small particles, such as microspheres or microrods that contain some method for identifying a set of particles composing one assay. Labeled particles have different target molecules that they assay for. These particles are added to a liquid (such as serum or cell lysate) containing the potential analytes. A system such as "smart dust" may use an alternative method of analyte signaling and readout.) The final step in the assay activates a reporter fluorophore that provides a signal. For each particle in the mixture, the cytometer identifies the classifier for the set the particle belongs to together with the fluorescence reading of the reporter fluorophore. Multi-well assay plates can be used to test many samples, and such assays become a high throughput system
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
