Sperm Inspection for In Vitro Fertilization via Self-Assembled Microdroplet Formation and Quantitative Phase Microscopy

We present a new method for the selection of individual sperm cells using a microfluidic device that automatically traps each cell in a separate microdroplet that then individually self-assembles with other microdroplets, permitting the controlled measurement of the cells using quantitative phase microscopy. Following cell trapping and droplet formation, we utilize quantitative phase microscopy integrated with bright-field imaging for individual sperm morphology and motility inspection. We then perform individual sperm selection using a single-cell micromanipulator, which is enhanced by the microdroplet-trapping procedure described above. This method can improve sperm selection for intracytoplasmic sperm injection, a common type of in vitro fertilization procedure.

Optical Fiber Tweezers for the Assembly of Living Photonic Probes

Optical fiber tweezers, as a versatile tool for optical trapping and manipulation, have attracted much attention in cell trapping, manipulation, and detection. Particularly, assembly of living cells using optical fiber tweezes has become a significant attention. Advanced achievements have been made on the assembly of fully biocompatible photonic probes with biological cells, enabling optical detection in biological environment in a highly compatible manner. Therefore, in this chapter, we discuss the use of optical fiber tweezers for assembly of living photonic probes. Living photonic probes can be assembled by the trapping and assembly of multiple cells using optical fiber tweezers. These photonic probes exhibit high biocompatibility and show great promise for the bio-applications in bio-microenvironments.

A Review of Advanced Impedance Biosensors with Microfluidic Chips for Single-Cell Analysis

Electrical impedance biosensors combined with microfluidic devices can be used to analyze fundamental biological processes for high-throughput analysis at the single-cell scale. These specialized analytical tools can determine the effectiveness and toxicity of drugs with high sensitivity and demonstrate biological functions on a single-cell scale. Because the various parameters of the cells can be measured depending on methods of single-cell trapping, technological development ultimately determine the efficiency and performance of the sensors. Identifying the latest trends in single-cell trapping technologies afford opportunities such as new structural design and combination with other technologies. This will lead to more advanced applications towards improving measurement sensitivity to the desired target. In this review, we examined the basic principles of impedance sensors and their applications in various biological fields. In the next step, we introduced the latest trend of microfluidic chip technology for trapping single cells and summarized the important findings on the characteristics of single cells in impedance biosensor systems that successfully trapped single cells. This is expected to be used as a leading technology in cell biology, pathology, and pharmacological fields, promoting the further understanding of complex functions and mechanisms within individual cells with numerous data sampling and accurate analysis capabilities.

Integrating magnetic capabilities to intracellular chips for cell trapping

Current microtechnologies have shown plenty of room inside a living cell for silicon chips. Microchips as barcodes, biochemical sensors, mechanical sensors and even electrical devices have been internalized into living cells without interfering their cell viability. However, these technologies lack from the ability to trap and preconcentrate cells in a specific region, which are prerequisites for cell separation, purification and posterior studies with enhanced sensitivity. Magnetic manipulation of microobjects, which allows a non-contacting method, has become an attractive and promising technique at small scales. Here, we show intracellular Ni-based chips with magnetic capabilities to allow cell enrichment. As a proof of concept of the potential to integrate multiple functionalities on a single device of this technique, we combine coding and magnetic manipulation capabilities in a single device. Devices were found to be internalized by HeLa cells without interfering in their viability. We demonstrated the tagging of a subpopulation of cells and their subsequent magnetic trapping with internalized barcodes subjected to a force up to 2.57 pN (for magnet-cells distance of 4.9 mm). The work opens the venue for future intracellular chips that integrate multiple functionalities with the magnetic manipulation of cells.

Engineered Membranes for Residual Cell Trapping on Microfluidic Blood Plasma Separation Systems. A Comparison between Porous and Nanofibrous Membranes

Blood-based clinical diagnostics require challenging limit-of-detection for low abundance, circulating molecules in plasma. Micro-scale blood plasma separation (BPS) has achieved remarkable results in terms of plasma yield or purity, but rarely achieving both at the same time. Here, we proposed the first use of electrospun polylactic-acid (PLA) membranes as filters to remove residual cell population from continuous hydrodynamic-BPS devices. The membranes hydrophilicity was improved by adopting a wet chemistry approach via surface aminolysis as demonstrated through Fourier Transform Infrared Spectroscopy and Water Contact Angle analysis. The usability of PLA-membranes was assessed through degradation measurements at extreme pH values. Plasma purity and hemolysis were evaluated on plasma samples with residual red blood cell content (1, 3, 5% hematocrit) corresponding to output from existing hydrodynamic BPS systems. Commercially available membranes for BPS were used as benchmark. Results highlighted that the electrospun membranes are suitable for downstream residual cell removal from blood, permitting the collection of up to 2 mL of pure and low-hemolyzed plasma. Fluorometric DNA quantification revealed that electrospun membranes did not significantly affect the concentration of circulating DNA. PLA-based electrospun membranes can be combined with hydrodynamic BPS in order to achieve high volume plasma separation at over 99% plasma purity.

A Microfluidic Array Device for Single Cell Capture and Intracellular Ca2+ Response Analysis Induced by Dynamic Biochemical Stimulus

A microfluidic array was constructed for trapping single cell and loading identical dynamic biochemical stimulation for gain a better understanding of Ca2+ signalling in single cells by applying extracellular dynamic biochemical stimulus. This microfluidic array consists of multiple radially aligned flow channels with equal intersection angles, which was designed by a combination of stagnation point flow and physical barrier. Numerical simulation results and trajectory analysis shown the effectiveness of this single cell trapping device. Fluorescent experiment results demonstrated the effects of flow rate and frequency of dynamic stimulus on the profiles of biochemical concentration which exposed on captured cells. In this array chip, the captured single cells in each trapping channels were able to receive identical extracellular dynamic biochemical stimuli which being transmitted from the entrance at the middle of the microfluidic array. Besides, after loading dynamic Adenosine Triphosphate (ATP) stimulation on captured cells by this device, consistent average intracellular Ca2+ dynamics phase and cellular heterogeneity were observed in captured single K562 cells. Furthermore, this device is able to be used for investigating cellular respond in single cells to temporally varying environments by modulating the stimulation signal in terms of concentration, pattern, and duration of exposure.

Topology Optimization of Passive Cell Traps

This paper discusses a flexible design method of cell traps based on the topology optimization of fluidic flows. Being different from the traditional method, this method obtains the periodic layout of the cell traps according to the cell trapping requirements by proposing a topology optimization model. Additionally, it satisfies the cell trapping function by restricting the flow distribution while taking into account the overall energy dissipation of the flow field. The dependence on the experience of the designer is reduced when this method is used to design a cell trap with acceptable trapping performance. By comparing the influence of the changes of various parameters on the optimization results, the flexibility of the topology optimization method for cell trap structure optimization is verified. The capability of this design method is validated by several performed comparisons between the obtained layouts and optimized designs in the published literature.