The Development of an Effective Bacterial Single-Cell Lysis Method Suitable for Whole Genome Amplification in Microfluidic Platforms

Single-cell sequencing is a powerful technology that provides the capability of analyzing a single cell within a population. This technology is mostly coupled with microfluidic systems for controlled cell manipulation and precise fluid handling to shed light on the genomes of a wide range of cells. So far, single-cell sequencing has been focused mostly on human cells due to the ease of lysing the cells for genome amplification. The major challenges that bacterial species pose to genome amplification from single cells include the rigid bacterial cell walls and the need for an effective lysis protocol compatible with microfluidic platforms. In this work, we present a lysis protocol that can be used to extract genomic DNA from both gram-positive and gram-negative species without interfering with the amplification chemistry. Corynebacterium glutamicum was chosen as a typical gram-positive model and Nostoc sp. as a gram-negative model due to major challenges reported in previous studies. Our protocol is based on thermal and chemical lysis. We consider 80% of single-cell replicates that lead to >5 ng DNA after amplification as successful attempts. The protocol was directly applied to Gloeocapsa sp. and the single cells of the eukaryotic Sphaerocystis sp. and achieved a 100% success rate

Point-of-Care Detection Devices for Healthcare

With recent technological advances in multiple research fields such as materials science, micro-/nano-technology, cellular and molecular biology, bioengineering and the environment, much attention is shifting toward the development of new detection tools that not only address needs for high sensitivity and specificity but fulfil economic, environmental, and rapid point-of-care needs for groups and individuals with constrained resources and, possibly, limited training. Miniaturized fluidics-based platforms that precisely manipulate tiny body fluid volumes can be used for medical, healthcare or even environmental (e.g., heavy metal detection) diagnosis in a rapid and accurate manner. These new detection technologies are potentially applicable to different healthcare or environmental issues, since they are disposable, inexpensive, portable, and easy to use for the detection of human diseases or environmental issues—especially when they are manufactured based on low-cost materials, such as paper. The topics in this book (original and review articles) would cover point-of-care detection devices, microfluidic or paper-based detection devices, new materials for making detection devices, and others