Microfluidics and Nanotechnology

7 Droplet-Based Digital Microfluidics for Single-Cell Genetic Analysis

Yong Zeng and Richard A. Mathies

CONTENTS

7.1 Introduction

7.2 High-Throughput, Programmable Microfluidic Droplet Generation

7.2.1 Experimental

7.2.1.1 MEGA Fabrication and Assembly

7.2.1.2 Droplet Generation

7.2.2 Results and Discussion

7.2.2.1 MEGA Design and Performance

7.2.2.2 Programmable Active Droplet Generation

7.3 Single-Cell Quantification and Genotyping of Bacterial Pathogens

7.3.1 Experimental

7.3.1.1 Cell Culture

7.3.1.2 Bead and PCR Preparation

7.3.1.3 PCR Reaction

7.3.1.4 Bead Recovery and Flow Cytometry Quantitation

7.3.2 Results

7.3.2.1 Digital Single-Cell Pathogen Detection at the Statistically Dilute Regime

7.3.2.2 High-Throughput and Sensitive SCGA