Microfluidic Cell Trapping for Single-Cell Analysis.

Bing Deng,Heyi Wang,Zhaoyi Tan,Yi Quan

doi:10.3390/mi10060409

Abstract

The single-cell capture microfluidic chip has many advantages, including low cost, high throughput, easy manufacturing, integration, non-toxicity and good stability. Because of these characteristics, the cell capture microfluidic chip is increasingly becoming an important carrier on the study of life science and pharmaceutical analysis. Important promises of single-cell analysis are the paring, fusion, disruption and analysis of intracellular components for capturing a single cell. The capture, which is based on the fluid dynamics method in the field of micro fluidic chips is an important way to achieve and realize the operations mentioned above. The aim of this study was to compare the ability of three fluid dynamics-based microfluidic chip structures to capture cells. The effects of cell growth and distribution after being captured by different structural chips and the subsequent observation and analysis of single cells on the chip were compared. It can be seen from the experimental results that the microfluidic chip structure most suitable for single-cell capture is a U-shaped structure. It enables single-cell capture as well as long-term continuous culture and the single-cell observation of captured cells. Compared to the U-shaped structure, the cells captured by the microcavity structure easily overlapped during the culture process and affected the subsequent analysis of single cells. The flow shortcut structure can also be used to capture and observe single cells, however, the shearing force of the fluid caused by the chip structure is likely to cause deformation of the cultured cells. By comparing the cell capture efficiency of the three chips, the reagent loss during the culture process and the cell growth state of the captured cells, we are provided with a theoretical support for the design of a single-cell capture microfluidic chip and a reference for the study of single-cell capture in the future.

Highlights

Mammalian eukaryotic cells generally are between 1–10 μm in diameter, and the content of a single cell is at the fL level [1]
With the deepening of the exploration of the laws of life, the demand for real-time and dynamic research methods has led to the emergence of new life analysis techniques and methods [2,3,4,5,6,7,8]
Lab-on-a-chip (LOC) was considered as a breakthrough technology, on the basis of its good manipulation of small volume liquids, such as cell isolation, localization and capture, to enable diverse cell-related studies at cellular, subcellular and molecular levels that can be performed at the micron scale

Summary

Introduction

Mammalian eukaryotic cells generally are between 1–10 μm in diameter, and the content of a single cell is at the fL level [1]. The realization of single-cell capture is the basis of cell culture, cell fusion, cell pairing and other operations, while a hydrodynamic microfluidic device can accurately control fluid flow, reduce cell culture time and cost, can provide cell analysis with the advantages of minimum dilution error, can achieve a sufficient number of individual cells to analyze the cell, can reveal the characteristics of individual cells and the differences between cells and can obtain more accurate and statistically significant data. The research results provide reference for different cell culture needs, and provide a reference for cell-level research, such as organ chip construction, analysis of single-cell stress response, interaction between cells and cells and for the development of radioactive cells This provides a reference for a more efficient and environmentally friendly research platform for the biological effects of experimental research

Cell Culture and Preparation

Cell Seeding

Chip Fabrications