Research on cells and organ‐like tissues is critical in the fields of molecular biology, genetic analysis, proteomics analysis, tissue engineering, and others. In recent years, advancements in precise cell manipulation technologies have made precise positioning and batch processing of cells feasible. Various methods are used for cell recognition, positioning, manipulation, and assembly, often introducing external fields such as electric, magnetic, acoustic, or optical fields into the liquid environment to interact with cells, applying forces to induce cell movement and rearrangement. Alternatively, three‐dimensional (3D) bioprinting technology is employed for precise cell positioning and assembly. This review will comprehensively assess the status, principles, advantages, disadvantages, and prospects of these precise cell manipulation technologies, covering single‐cell manipulation, multicellular assembly, and biological 3D printing techniques.

The heterogeneity of circulating tumor cells has a significant impact on the diagnosis, treatment, and monitoring of cancer. Research on the subtypes of circulating tumor cells can bring better treatment outcomes for cancer patients. Here, we proposed a microfluidic chip for the magnetic capture of subtypes of circulating tumor cells from the whole blood and phenotypic profiling by stacking laminar flow vertically. Circulating tumor cells were sorted and captured by the three-dimensional regulation of both magnetic fields in the vertical direction and flow fields in the lateral direction. Using EpCAM-magnetic beads, we achieved sorting and sectional capture of target cells in whole blood and analyzed the surface expression levels of the captured cells, confirming the functionality of the microfluidic chip in sorting and capturing subtypes of circulating tumor cells. This microfluidic chip can also aid in the subsequent subtype analysis of other rare cells.

Heterogeneous expression of a single surface protein within one cell population can drive major functional differences, yet low-expression subtypes remain difficult to isolate. Conventional tube-based immunomagnetic separation collapses all labelled cells into one positive fraction and thus cannot resolve small differences in marker abundance. Here, we present MagSculptor, a microfluidic platform for high-resolution magnetic fractionation of low-expression EpCAM-defined subtypes within one immunomagnetically labelled population at a time. Arrays of soft-magnetic strips create localized high-gradient zones that map modest differences in bead loading onto distinct capture positions, yielding High (H), Medium (M), Low (L), and Negative (N) fractions. Finite element method simulations of coupled magnetic and hydrodynamic fields quantify the field gradients and define an operating window. Experimentally, epithelial cancer cell lines processed sequentially under identical settings show reproducible subtype partitioning. In a low-EpCAM model (MDA-MB-231), conventional flow cytometry, under standard EpCAM staining conditions, did not yield a robust EpCAM-positive gate, whereas MagSculptor still revealed graded subpopulations. Western blotting confirms a monotonic decrease in EpCAM abundance from H to N, and doxorubicin assays show distinct in vitro drug sensitivities, while viability remains above 95%. MagSculptor thus helps extend immunomagnetic separation from binary enrichment to multi-level isolation of low-expression subtypes and provides a convenient front-end for downstream functional and molecular analyses.

Continuous-flow separation of magnetically labeled cells according to surface-marker expression levels is increasingly needed to study phenotypic heterogeneity and support downstream assays. Here, we present a microfluidic platform that uses spatially engineered soft magnetic strips (SMS) to sculpt lateral magnetic deflection fields for quantitative, label-guided cell fractionation. Under a uniform bias field, the SMS generates controllable magnetic gradients within the microchannel, producing distinct lateral velocities among EpCAM-labeled tumor cells that carry different Dynabead loads, which indirectly report membrane protein expression. Multi-outlet collection converts these “race-based” trajectory differences into discrete expression-level-resolved fractions. A COMSOL–MATLAB framework and a force-equivalent metric |(H·∇)H| are used to optimize key structural parameters of the magnetic interface, including strip thickness, width, and vertical spacing from the flow channel. Three journey nodes at 1.5, 3, and 9 mm along the flow path define a three-stage cascade that partitions MDA-MB-231, Caco-2, and A549 cells into four EpCAM-related magnetic subgroups: high (H), medium (M), low (L), and near-negative (N). Experiments show that the sorted fractions follow the expected expression trends reported in the literature, while maintaining high cell recovery (>90%) and viability retention of 98.2 ± 1.3%, indicating compatibility with downstream whole-blood assays and culture. Rather than introducing a new biomarker, this work establishes a quantitative magnetic-field design strategy for continuous microfluidic sorting, in which the spatial configuration of soft magnetic elements is exploited to implement expression-level-dependent fractionation in next-generation magneto-fluidic separation systems.

Subtype isolation of cells with low surface protein expression remains a longstanding bottleneck in various biological and biomedical investigations. Herein, we present MagSculptor, a novel bio-magnetic separator that leverages passive magnetic field shaping through engineered soft magnetic strip arrays embedded within a microfluidic platform. By integrating high-permeability amorphous soft magnetic materials with a tunable background magnetic field, the system creates a series of localized high-flux density zones with magnetic field gradients exceeding 2,000 T/m at a distance of 50 μm. Both numerical simulations and experimental results demonstrate that MagSculptor enables dynamic and highly resolute separation of magnetically labeled cells by precisely modulating the external magnetic field intensity and the spatial arrangement of soft magnetic elements. Consequently, cells can be classified into four distinct subtypes: High (H), Medium (M), Low (L), and Negative (N), based on magnetic labeling, which correlates linearly with the target protein expression level. The system exhibited superior resolution in sorting cancer cell lines such as MDA-MB-231 and A549 based on EpCAM expression, with cell viability exceeding 95%, making it a powerful and versatile tool for downstream applications in single-cell transcriptomics, genomics, and mechanistic biological investigations.