Multiplexed Quantification
Multiplexed Quantification - Related Videos
00:07
Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation
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Cited by 14 •
2019
Quantitative Multiplex Immunoprecipitation (QMI) uses flow cytometry for sensitive detection of differences in the abundance of targeted protein-protein interactions between two samples. QMI can be performed using a small amount of biomaterial, does not require genetically engineered tags, and can be adapted for any previously defined protein interaction network.
09:38
Multiplexed Immunofluorescence Assay for Spatial Assessment of Senescence Markers in vivo
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2026
Multiplexed immunofluorescence enables sequential labeling of multiple antibody markers within a single tissue section to support spatial analysis of tissue architecture and cell populations. Here, a workflow for multiplex detection of senescence-associated, structural, and immune markers is presented, including manual and automated slide preparation, iterative imaging, and single-cell analysis using digital pathology software.
13:14
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Cited by 4 •
2015
Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.
08:05
Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
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Cited by 19 •
2011
Using Luminex Corporation’s xMAP microsphere technology, we have developed the Multiplexed Fluorometric ImmunoAssay (MFIA) for serosurveillance of various laboratory animal species. The MFIA is a suspension microarray where antigen, tissue control or immunoglobulins are covalently linked to color-coded polystyrene microspheres. The MFIA testing method as well as various troubleshooting topics is addressed.
05:22
A Multiplex Serological Assay for the Detection of Antibody Responses to Arboviruses
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2025
Multiplex fluorescence immunoassays are becoming a reference for serology testing. Their application, however, is limited by the complexity and time-consuming bead-analyte coupling processes. This report presents a semi-automated, reproducible multiplex microsphere bead coupling protocol that is comparable to the standard method while considerably reducing time to results.
FAQs
Each analyte or biological target receives a distinguishable signal, which may come from a fluorescent label, antibody, nucleic acid probe, or molecular barcode. The measurement system then separates the signals and calibrates them so that each signal can be assigned to its corresponding target. This enables several measurements from the same sample without treating all detected signals as one combined result.
Calibration connects the strength of a detected signal with the concentration of its corresponding analyte. Without this step, the presence of several signals would indicate that targets were detected but would not support reliable individual quantification. Calibration therefore allows researchers to compare cytokine, antibody, or pathogen-marker levels within a sample and across disease or treatment conditions.
The approach can use fluorescent labels, antibodies, nucleic acid probes, or molecular barcodes to distinguish targets. These signal systems can be applied to different biological measurements, including cytokines, antibodies, pathogen markers, and cellular responses. The choice of signal determines how targets are identified and separated during analysis, while calibration supports estimation of their individual concentrations.
A study first identifies the analytes or biological targets relevant to its question, then assigns each target a distinguishable signal. The sample is measured for all selected signals, after which the signals are separated and calibrated to determine target-specific concentrations. This workflow converts a complex measurement from one sample into results that can be examined target by target.
It is useful when investigators need to examine several parts of an immune or infectious process from limited sample volume. A single analysis can include cytokines, antibodies, pathogen markers, and cellular responses, helping characterize host-pathogen interactions. It can also support comparisons between disease states and assessments of therapeutic or vaccine responses.
The combined measurements can reveal patterns across immune and infectious targets rather than focusing on one parameter alone. In immunology and infection research, these datasets help characterize host-pathogen interactions, identify potential biomarkers, compare disease states, and evaluate responses to therapies or vaccines. Their value comes from relating multiple quantified signals within the same biological context.