Multiplexed flow cytometry, known for its powerful high-throughput
identification capability, is widely applied across various biomedical
and clinical fields. However, classical flow cytometry relies on multichannel
lasers and detectors, which are significant in cost and size, limiting
their application in miniaturized assays. Herein, we developed an
acoustic streaming-based flow cytometry technique that focuses on
multisized microbeads flowing sheathlessly. This method enables the
discrimination of particle types and the quantification of target
protein concentrations using only a single detector. Microbeads of
different sizes exhibit distinct behaviors in the continuous acoustic
streaming tunnel, leading to an increased velocity difference during
their transition under the laser spot. Consequently, a size detection
method based on “velocity stretching” has been established.
A multiplex assay of three proteins: cardiac troponin I, creatine
kinase-MB and myoglobin, in acute myocardial infarction is performed
to validate the feasibility and evaluate the performance of the system.
This new multiplexed flow cytometry strategy is expected to enable
low-cost and onsite detection of multiple biomarkers.