研究目的
To develop a label-free and non-invasive method for precise separation of leukocyte subpopulations (lymphocytes, monocytes, granulocytes) using a hybrid acoustic-optical chip for applications in leukemia diagnosis and analysis.
研究成果
The hybrid acoustic-optical chip enables precise, label-free separation of leukocyte subpopulations with high purity (up to 99% for lymphocytes, 98% for monocytes, 95% for granulocytes) and good cell viability. This non-invasive method overcomes limitations of traditional techniques and shows promise for clinical applications in leukemia diagnosis and analysis, with potential for future improvements in rare cell detection.
研究不足
The method may have limitations in throughput compared to some high-throughput systems, and the complexity of device fabrication could be a barrier. Optimization of parameters like flow rate and laser power is necessary to maintain high purity, and the technique may require further validation with diverse clinical samples.
1：Experimental Design and Method Selection:
The study integrates acoustic and optical technologies in a microfluidic chip. Standing surface acoustic waves (SSAW) are used for 3D focusing and initial separation based on size, while optical forces are applied for precise separation based on refractive index differences. Theoretical models include equations for acoustic radiation force and optical force calculations.
2：Sample Selection and Data Sources:
Human peripheral blood samples were obtained with informed consent. Peripheral blood mononuclear cells (PBMCs) and granulocytes were isolated using discontinuous gradient centrifugation. Samples were cultured and maintained in RPMI 1640 medium with fetal bovine serum.
3：List of Experimental Equipment and Materials:
Microfluidic chip fabricated with PDMS on a lithium niobate substrate, interdigital transducers (IDTs), laser source (1064 nm wavelength), confocal microscope (Nikon A1R), high-speed camera (Hamamatsu ORCA-Flash 4.0 v2), fluorescence microscope (Olympus IX81), cell counter (Nexcelom Cellometer Mini), photoresist (SU-8), antibodies for staining (e.g., FITC-labeled anti-CD3, Cy5-labeled anti-CD14).
4：0 v2), fluorescence microscope (Olympus IX81), cell counter (Nexcelom Cellometer Mini), photoresist (SU-8), antibodies for staining (e.g., FITC-labeled anti-CD3, Cy5-labeled anti-CD14).
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: The micro-channel was fabricated using soft lithography. IDTs were patterned on the substrate. Blood samples were prepared and stained for visualization. Acoustic waves were generated by applying RF signals to IDTs for focusing and separation. Optical separation was performed using a laser beam. Cell trajectories and separation efficiency were monitored and recorded using microscopy and high-speed imaging.
5：Data Analysis Methods:
Separation purity and recovery rates were calculated based on cell counts from outlet samples. Statistical analysis included standard deviations from multiple tests. Simulations of acoustic and optical forces were conducted using theoretical equations.
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