Abstract
Multiplexing allows quantifying multiple analytes in a single step, providing advantages over individual testing through shorter processing time, lower sample volume, and reduced cost per test. Currently, flow cytometry is the gold standard for biomedical multiplexing, but requires technical training, extensive data processing, and expensive operational and capital costs. To solve this challenge, we designed digital barcoded particles and a microfluidic architecture for multiplexed analyte quantification. In this work, we simulate and model non-fluorescence-based microfluidic impedance detection with a single excitation and detection scheme using barcoded polymer microparticles. Our barcoded particles can be designed with specific coding regions and generate numerous distinct patterns enabling digital barcoding. We found that signals based on adhered microsphere position and relative orientation were evaluated and separated based on their associated electrical signatures and had a 7 µm microsphere limit of detection. Our proposed microfluidic system can enumerate micron-sized spheres in a single assay using barcoded particles of various configurations. As representation of blood cells, the microsphere concentrations may provide useful information on disease onset and progression. Such sensors may be used for diagnostic and management of common critical care diseases like sepsis, acute kidney injury, urinary tract infections, and HIV/AIDS.
Highlights
Most diagnostic equipment requires large blood volumes to measure each targeted analyte separately
Flow cytometry is employed for obtaining heterogenous cell sample characteristics beyond a CBC as a mainstream multiplexing system
Flow cytometry is ubiquitously used as the diagnosing standard, it has severe shortcomings in cost, unable to quantify secreted or dissolved compounds, complicated data analysis, and poor receiver operating curves for in-the-field use[2,7,14,15,16,17,18]
Summary
Most diagnostic equipment requires large blood volumes to measure each targeted analyte separately. Several researchers are pioneering miniaturized, novel methods as point-of-care alternatives to flow cytometry, including fluorescent microfluidics for nucleic acid and protein quantification[19,20,21], droplet-based microfluidics for secreted biomarker analysis[17,22,23], planar microarrays with micro-engraving techniques for temporal cell behavior evaluation[24,25,26,27], and barcoded microchip devices for membrane and cytosolic protein detection[28,29,30] Such proceedings can be directed for treating complicated and changing diseases requiring multiple biomarker determinations like sepsis[5,7,15,31,32], HIV/AIDS9,33–35, acute kidney injury[36,37,38], urinary tract infections[12,22], and malignant tumors[26,30,39,40,41]. We evaluate the design and simulate impedance-based microfluidic behaviors with novel barcoded particles for cell surface receptor detection
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