Node-pore sensing enables label-free surface-marker profiling of single cells.

Area of Science:

• Biotechnology
• Cell Biology
• Microfluidics

Background:

• Traditional flow cytometry is a multiparametric method for cellular characterization.
• Limitations include spectral overlap, compensation, lengthy preparation, and high costs, restricting its use to core facilities.
• There is a need for simpler, more affordable cellular screening technologies.

Purpose of the Study:

• To develop a low-cost, label-free microfluidic method for single-cell phenotypic profiling.
• To demonstrate the method's capability in analyzing cell lines and clinical samples.
• To offer a user-friendly and sensitive alternative to conventional flow cytometry.

Main Methods:

• Development of a microfluidic channel functionalized with antibodies for specific cell-surface antigens.
• Utilized Node-Pore Sensing to measure single-cell transit times as an indicator of antigen presence.
• Applied the method to screen acute promyelocytic leukemia cell lines and primary acute myeloid leukemia bone marrow samples.

Main Results:

• Simultaneously screened NB4 and AP-1060 cell lines for myeloid antigens (CD13, CD14, CD15, CD33).
• Distinguished between AP-1060 and NALM-1 cells based on CD13 and HLA-DR expression.
• Successfully identified and screened blast populations in primary acute myeloid leukemia bone marrow samples for CD13, CD34, and HLA-DR.

Conclusions:

• The label-free microfluidic Node-Pore Sensing method provides an affordable, sensitive, and user-friendly approach to cellular screening.
• This technology has the potential to simplify and decentralize cellular analysis, moving it from core facilities to benchside applications.
• The method effectively characterizes cellular phenotypes in both cell lines and complex clinical samples.